Selenophene anti-tumor agents

ABSTRACT

Novel selenophene compounds useful as anti-tumor agents are described. Preferred compounds include compounds of formula I:  
                 
 
     wherein R 1  and R 2  are independently selected from the group consisting of;  
                 
 
      H, CHO, CH 2 OH and CH 2 NH 2 ; and  
     X and Y are independently selected from the group consisting of Se, S, O, NCH 3  and NH. Pharmaceutical compositions and a method for treating patients having tumors utilizing the disclosed selenophene compounds are also described.

GOVERNMENT RIGHTS

[0001] This invention was made with United States Government supportunder Grant No. UO1 CA50743, awarded by the National Cancer Institute.The United States Government has certain rights in the invention.

FIELD OF THE INVENTION

[0002] The present invention relates to compositions and a method fortreating a patient having a tumor. More specifically, the presentinvention relates to the treatment of such patients with an effectiveamount of a selenophene derivative.

BACKGROUND AND SUMMARY OF THE INVENTION

[0003] The control and cure of cancer represents one of our mostchallenging health problems. The treatment of cancer can be approachedby several modes of therapy including surgery, radiation, chemotherapyor a combination of any of these treatments. Chemotherapy continues tobe an indispensable therapy for inoperable or metastatic forms of thedisease.

[0004] The selection of natural compounds, or the synthesis of newcompounds having effective anticancer activity is complicated by thestill limited knowledge of cancer cell biology and biochemistry.Therefore, development of new effective anti-tumor agents will remainheavily dependent on screening compounds to discover novel compoundshaving cytotoxic activity. Preferably, such compounds exhibit enhancedcytotoxicity against tumor cells relative to their cytotoxicity tonormal cells.

[0005] The success of novel antitumor drug development programs isdependent on the initial identification of antitumor agents. Thus thediscovery of antitumor agents requires the systematic screening of alarge number of natural products and synthetic compounds.

[0006] The mouse L1210 leukemia cell line was initially the preferredmodel system used for screening natural compounds for antitumoractivity. However, the P388 murine leukemia system was found to be moresensitive and predictive than L1210 leukemia system, and has been usedas primary screen during the past decade. Systematic screening forcompounds exhibiting toxicity to these two leukemia cell lines hasresulted in the isolation of a large number of active natural products.However, the anticancer activities of these compounds were predominantlyin leukemia, lymphoma and a few rare tumors. Low clinical efficacy, orthe lack of clinical efficacy of known chemotherapeutics against slowergrowing solid tumors, is a serious concern.

[0007] It has been recognized that the use of a single antileukemiascreening system could bias the end results and lead to the isolation ofcompounds only active in the treatment of fast growing tumors. Inaddition, the use of a single antileukemia screening system may notdetect novel compounds with high specificities for particular celllines. It is also likely that many novel compounds with possibleanti-tumor activity have remained undetected by the less sensitive invivo models due to the low concentrations at which many active naturalproducts occur.

[0008] Considering the diversity of tumors in terms of cell type,morphology, growth rate and other cellular characteristics, the U.S.National Cancer Institute (NCI) has developed a “disease-oriented”approach to antitumor activity screening (M. R. Boyd, in “Principle ofPractice of Oncology” J. T. Devita, S. Hellman, S. A. Rosenberg (Eds.)Vol. 3, PPO Update, No. 10, 1989). This in vitro prescreening system isbased on the measurement of antitumor cytotoxicity against human tumorcell line panels consisting of approximately 60 cell lines of majorhuman tumors (including leukemia and slower growing tumor cells such aslung, colon, breast, skin, kidney, etc.). The most important advantageof the new in vitro screening panels is the opportunity to identifycompounds that are selectively more cytotoxic to cells of slowly growingsolid tumors than to rapidly growing leukemia cells.

[0009] The cytotoxicity profile of the NCI human tumor cell panelsdisplays the tumor specificity of a given compound, however the assaydoes not assess the toxicity of that compound to normal human cells.Accordingly a second bioassay is utilized to measure the selectivecytotoxicity against certain types of tumor cells verses normal humancells.

[0010] The growth and differentiation of cells are regulated bysignaling cascades induced by various mitogenic proteins (J. Kurjan andB. L. Taylor, “Signal Transduction,” Academic Press, New York, N.Y.1994) that often are encoded by proto-oncogenes. The overexpression,amplification or mutation of the oncoprotein is critically involved inthe initiation, progression and metastasis of malignant cells (R. A.Weinberg, “Oncogenes and the Molecular Origins of Cancer,” Cold SpringHarbor Laboratory Press, Cold Spring Harbor, N.Y., 1989). Manyoncoproteins alter normal cellular growth regulation by modulating theintracellular signaling pathways from the membrane to the nucleus.Therefore, cancer may be considered as a disease of cellular signaltransduction, which presents a novel approach for anticancer therapy.One of the critical enzymes involved in the oncoprotein signaltransduction is protein kinase C (U. Nishizuka, Nature, 308, 693, 1984and Science, 233, 305, 1986). Thus, the determination of a compound'sability to inhibit protein kinase C activity has become a goodprognostic for discovering novel anticancer agents (A. Basu, PharmacTher, 59, 257, 1993). Furthermore it is anticipated that the selenophenecompounds will demonstrate selectivity for certain class members ofprotein kinases, including protein kinase C. Inhibition of a specificclasses of protein kinases will allow the treatment of other diseasesassociated with defects in signaling transduction.

[0011] Selenophenes are selenium containing heterocyclic compounds thatare analogs of naturally occurring thiophene, furan and pyrrolecompounds. Selenophenes have been found to be effective antitumoragents, and exhibit enhanced cytotoxicity against slow growing tumorcells, selective cytotoxicity against human renal, ovarian tumor cells,and skin tumor cells; and exhibit inhibition of protein kinase C.

[0012] In accordance with this invention there is provided a method forthe treatment of cancer which utilizes selenophene compounds of theformula I:

[0013] wherein R₁ and R₂ are independently selected from the groupconsisting of;

[0014]  H, CHO, CH₂OH and CH₂NH₂;

[0015] X and Y are independently selected from the group consisting ofSe, S, O, NCH₃ and NH;

[0016] R₃, R₄, R₅ and R₆ are independently selected from the groupconsisting of H, CHO, CH₂OH and CH₂NH₂; cyclodextrin complexes of suchcompounds; and when R₃, R₄, R₅ or R₆ is CH₂NH₂, the pharmaceuticallyacceptable salt of the compound represented thereby.

[0017] Further in accordance with this invention there are providednovel cytotoxic compounds of the above formula and chemotherapeuticpharmaceutical compositions containing said compounds in anti-tumoreffective amounts.

[0018] Additional objects, features, and advantages of the inventionwill become apparent to those skilled in the art upon consideration ofthe following detailed description of preferred embodiments exemplifyingthe best mode of the invention as presently perceived.

DETAILED DESCRIPTION OF THE INVENTION

[0019] The present invention is directed to selenophene compounds, theirpharmaceutical compositions and methods utilizing suchcompounds/compositions for treating patients, having tumor. Theselenophene compounds are effective antitumor agents against slowgrowing tumors, and generally have been found to exhibit high selectivecytotoxicity for individual tumor cell lines.

[0020] The compounds of the present invention are selenophene compoundsof the formula I:

[0021] wherein R₁ and R₂ are independently selected from the groupconsisting of;

[0022]  H, CHO, CH₂OH and CH₂NH₂;

[0023] X and Y are independently selected from the group consisting ofSe, S, O and NR;

[0024] R is H or C₁-C₇ alkyl;

[0025] R₃, R₄, R₅ and R₆ are independently selected from the groupconsisting of nitro, amino, alkoxy, cyano, chloro, bromo, iodo, C₁-C₇alkyl or haloalkyl, C₁-C₇ alkenyl or haloalkenyl, C₁-C₄ alkanoyloxymethyl, CH₂OR₇, COR₈, CH₂NR₉R₁₀, CH(OR₇)R₁₁, CH═CR₁₂R₁₃, CH═NR₁₄,CH₂SC(NH)NH₂ and C≡CR₁₅ wherein

[0026] R₇ is H, CO(CH₂)₂CO₂H, (CH₂)₂OCH₃, C₁-C₄ alkyl or COC₁-C₁, alkyl;

[0027] R₈ is H or C₁-C₇ alkyl;

[0028] R₉ and R₁₀ are independently H, CN, C₁-C₄ alkyl, or mono- ordi-hydroxyC₂-C₄ alkyl;

[0029] R₁₁ is C₁-C₇ alkyl, or C₁-C₇ alkenyl;

[0030] R₁₂ and R₁₃ are independently H, C₁-C₇ alkyl, COOR₈, CN,CH(OR₇)COOR₈, Br, CO-thienyl, COC₆H₄OH(p);

[0031] R₁₄ is NHR₇ or OR₈;

[0032] R₁₄ is COOR₈, CH(OR₇)CH₂OR₁₆ or CH(OCOC₁-C₄ alkyl)CH₂OR₈;

[0033] R₁₆ is H, COCH₂CH₂CO₂H, or COC₁-C₇ alkyl;

[0034]  cyclodextrin complexes of such compound and

[0035] when R₃, R₄, R₅ or R₆ is CH₂NR₆R₇, the pharmaceuticallyacceptable salt of the compound represented thereby.

[0036] In one preferred embodiment of this invention there is providedanti-tumor selenophenes of the above formula I,

[0037] wherein R₂ is

[0038] X and Y are independently selected from the group consisting ofS, Se and NH;

[0039] R₁ R₃, and R₆ are H; and

[0040] R₅ is selected from the group consisting of CHO or CH₂OH; andcyclodextrin complexes of such compounds. These compounds have beendemonstrated to exhibit cytotoxic selectivity against transformed humancells (See Table 1).

[0041] In another preferred embodiment of this invention there isprovided anti-tumor selenophenes of the above formula I wherein R₁ is

[0042] X and Y are independently selected from the group consisting ofS, Se and NH;

[0043] R₂, R₃, and R₆ are H;

[0044] R₅ is selected from the group consisting of CHO or CH₂OH; andcyclodextrin complexes of such compounds.

[0045] Other preferred compounds in accordance with this invention areselenophenes of formula I:

[0046] wherein R₁ and R₂ are independently selected from the groupconsisting of;

[0047] H, CHO, CH₂OH and CH₂NH₂;

[0048] X and Y are independently selected from the group consisting ofSe, S, O, NCH₃, and NH;

[0049] R₃, R₄, R₅ and R₆ are independently selected from the groupconsisting of H, CHO, CH₂OH and CH₂NH₂; cyclodextrin complexes of suchcompounds; and when R₂ or R₃ is CH₂NH₂, the pharmaceutically acceptablesalt of the compound represented thereby; with the proviso, that when R₂is

[0050] R₁ is other than

[0051] and when R₁ is

[0052] R₂ is other than

[0053] In accordance with another embodiment of the present inventionnovel intermediates of Formula II are also provided:

[0054] wherein W is selected from the group consisting of N(CH₃)₂ and

[0055] and X and Y are independently selected from the group consistingof Se, S, O, NCH₃ and NH.

[0056] One aspect of the present invention is a method of preparing thecompounds of Formula I through an intermediate a compound of theformula:

[0057] in accordance with the general methods of schemes 1-4 asdescribed hereinbelow, wherein X and Y are independently selected fromthe group consisting of Se, S, O, NCH₃ and NH.

[0058] The selenophene compounds of this invention are readilyformulated into pharmaceutical compositions, also within the scope ofthis invention, for use in the presently described method for treatmentof patients having tumors. In one preferred embodiment of thisinvention, the pharmaceutical composition comprises an anti-tumoreffective amount of a selenophene compound of formula I:

[0059] wherein R₁ and R₂ are independently selected from the groupconsisting of;

[0060]  H, CHO, CH₂OH and CH₂NH₂;

[0061] X and Y are independently selected from the group consisting ofSe, S, O and NR, wherein R is H or C₁-C₇ alkyl;

[0062] R₃, R₄, R₅ and R₆ are independently selected from the groupconsisting of nitro, amino, alkoxy, cyano, chloro, bromo, iodo, C₁-C₇alkyl or haloalkyl, C₁-C₇ alkenyl or haloalkenyl, C₁-C₄ alkanoyloxymethyl, CH₂OR₇, COR₈, CH₂NR₉R₁₀, CH(OR₇)R₁₁, CH═CR₁₂R₁₃, qH═NR₁₄,CH₂SC(NH)NH₂ and C≡CR₁₅ wherein

[0063] R₇ is H, CO(CH₂)₂CO₂H, (CH₂)₂OCH₃, C₁-C₄ alkyl or COC₁-C₁₇ alkyl;

[0064] R₈ is H or C₁-C₇ alkyl;

[0065] R₉ and R₁₀ are independently H, CN, C₁-C₄ alkyl, or mono- ordi-hydroxyC₂-C₄ alkyl;

[0066] R₁₁ is C₁-C₇ alkyl, or C₁-C₇ alkenyl;

[0067] R₁₂ and R₁₃ are independently H, C₁-C₇ alkyl, COOR₈, CN,CH(OR₇)COOR₈, Br, CO-thienyl, COC₆H₄OH(p);

[0068] R₁₄ is NHR₇ or OR₈;

[0069] R₁₅ is COOR₈, CH(OR₇)CH₂OR₁₆ or CH(OCOC₁-C₄ alkyl)CH₂OR₈;

[0070] R₁₆ is H, COCH₂CH₂CO₂H, or COC₁-C₇ alkyl;

[0071]  cyclodextrin complexes of such compound and

[0072] when R₃, R₄, R₅ or R₆ is CH₂NR₆R₇, the pharmaceuticallyacceptable salt of the compound represented thereby, and apharmaceutically acceptable carrier.

[0073] Another pharmaceutical composition within the scope of thisinvention comprises an anti-tumor effective amount of a selenophenecompound of the formula I:

[0074] wherein R₁ and R₂ are independently selected from the groupconsisting of;

[0075] H, CHO, CH₂OH and CH₂NH₂;

[0076] X and Y are independently selected from the group consisting ofSe, S, O, NCH₃ and NH;

[0077] R₃, R₄ and R₆ are H;

[0078] R₅ is selected from the group consisting of H, CHO, CH₂OH andCH₂NH₂; cyclodextrin complexes of such compounds; and when R₃, R₄, R₅ orR₆ is CH₂NH₂, the pharmaceutically acceptable salt of the compoundrepresented thereby; with the proviso, that when R₂ is

[0079]  R₁ is other than

[0080]  and when R₁ is

[0081]  R₂ is other than

[0082] and a pharmaceutically acceptable carrier.

[0083] The present compounds are readily prepared using art-recognizedchemical-synthesis procedures as exemplified hereinbelow.

[0084] The cytotoxic activity of the present selenophene compounds havebeen measured utilizing two different assays or screens. The firstscreen measures the cytotoxicity against a, panel of sixty differenthuman tumor cell lines. This assay provides data regarding the generalcytotoxicity of an individual compound. In particular this type of assayis useful in identifying compounds which have enhanced cytotoxicactivity against slow growing tumors as compared to faster growing tumorcells such as leukemia tumor cell lines. The identification of suchcompounds is critical since previously identified antitumor agents havelow cytotoxic activity against slower growing tumors. The specificity ofa compound for a limited number of tumor cell lines also indicates thatsuch a compound will likely be less cytotoxic to normal cells. Thespecificity of a cytotoxic compound for tumor cell lines relative tonormal cells is an important characteristic of an effective antitumoragent.

[0085] Antitumor cytotoxicity data generated from the National CancerInstitute human tumor cell panels can also be expressed in a graphicpattern (mean graph) to display differential cell growth inhibition (K.D. Paull, R. H. Shoemaker, L. Hodes, A. Monks, D. A. Scudiero, L.Rubinstein, J. Plowman and M. R. Boyd, J. Natl. Cancer Inst., 81, 1088,1989.) In the mean graph, the arithmetic mean of the logarithm of theGI₅₀ (50% growth inhibition), TGI (total growth inhibition) or LC₅₀ (50%lethal concentration) values is used as an anchor point. Relativecytotoxicity is displayed by projecting bars to the right or left of themean, depending on whether cell sensitivity to a test compound is moreor less than average. The length of a bar is indicative of differentialcytotoxicity against a specific type of tumor cells or tumor panels.

[0086] In a second assay, the cytotoxic selectivity is assessed bycomparing compound cytotoxicity against human renal carcinoma cells(A-498), ras-transformed human bronchial epithelial cells (TBE) andnormal human renal cells (RPTEC). IC₅₀ values were compared betweentreated TBE cells and RPTEC cells and the selective cytotoxicity index(SCI) was determined [SCI=GI₅₀(RPTEC)/GI₅₀(A-498)].

[0087] The antitumor cytotoxicity of the selenophene compounds tested inthose in vitro assays was measured by a microculture assay using eithera 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) orsulforhodamine B (SRB) based assay. [M. R. Boyd in “Principles andPractices of Oncology,” V. T. DeVita, Jr.]. The experiments wereconducted at Purdue University in 96-well microtiter plates and thecytotoxic effects of the selenophene compounds on those cells weremeasured by cell count using a Coulter Z. F. counter (Hialeah, Fla.).The results are expressed as GI₅₀, the concentration of drug at whichcell numbers are reduced to 50% of control cell culture [T. C. K. Chan,C. J. Chang, N. M. Koonchanok and R. L. Geahlen, Biochem. Biophys. Res.Commun., 193, 1152, (1993); S. Hellman and S. A. Rosenberg (Eds.), Vol.3, PPO Updates, Number 10, (1989).]

[0088] This in vitro microculture assay has an advantage over in vivoassays in that results are obtained within a week as opposed to severalmonths. The MTT assay is based on the production of a dark blue formazanproduct by dehydrogenase in the mitochondria of live tumor cells afterexposure to drug for 6 days [M. C. Alley, D. A. Scudiero, A. Monks, M.L. Hursey, M. J. Czerwinski, D. L. Fine, B. J. Abbott, J. G. Mayo, R. H.Shoemaker and M. R. Boyd, Cancer Res., 48, 589, 1988]. Thus, only livecells are stained and can be measured at 570 nm. The SRB assay is basedon the binding of the anionic group to the basic amino acid residues ofcellular proteins after exposure of tumor cells to drug for 2 days [P.Skehan, R. Storeng, D. Scudiero, A. Monks, J. McMahon, D. Vistica, J. T.Warren, H. Bohesch, S. Kenney and M. R. Boyd, J. Nat. Cancer Inst., 82,1107, 1990.] Thus, the total protein can be measured at 564 nm.Antitumor cytotoxicity is reported as GI₅₀, effect drug dose at whichcell growth is retarded to 50% of control culture of tumor cells. Theactive compounds are defined as those compounds having GI₅₀ values thatare less, than 10⁻⁴ M or 10 μg/ml.

[0089] The data presented in Table 1 illustrates that selenophenesgenerally exhibit greater cytotoxicity for human renal carcinoma cellsin comparison to the normal human cells. The data of Table 1demonstrates the selective cytotoxicity of various selenophene compoundsagainst human renal carcinoma and ras-oncogene transformed humanbronchial epithelial cells [in GI₅₀(ug/ml)]. The following abbreviationsare used for the tested cell lines. TABLE 1 NSC GI₅₀ (μg/ml) NumberRPTEC A-498 TBE SCI 674973 4 × 10⁰ 3 × 10⁰ 4 × 10⁰   1 675246 1 × 10⁻¹ 3× 10⁻⁶ 3 × 10⁻³ >1000 3 × 10⁻² 3 × 10⁻⁶ 2 × 10⁻³ >1000 675247 2 × 10⁻¹ 7× 10⁻⁵ 3 × 10¹ >1000 8 × 10⁰ 2 × 10⁻⁶ 2 × 10¹ >1000 676628 4 × 10² 8 ×10¹ 1 × 10²   5 676632 2 × 10⁻³ 3 × 10⁻⁷ <10⁻³ >1000 3 × 10⁻⁴ 2 × 10⁻⁷ 2× 10⁻⁴ >1000 675347 2 × 10¹ 3 × 10¹ 1 × 10¹ <1 675344 <10⁻² 3 × 10⁻⁷<10⁻² >1000 1 × 10⁻⁴ 6 × 10⁻⁸ 7 × 10⁻⁶ >1000 676633 2 × 10¹ 1 × 10² 1 ×10¹ <1 676634 1 × 10¹ 6 × 10⁻⁴ 3 × 10⁻⁴ >1000 4 × 10⁻¹ 3 × 10⁻³ 6 ×10⁻³ >100 676635 2 × 10⁰ <10⁻³ 2 × 10¹ >1000 123127 5 × 10⁻² 5 × 10⁻² 3× 10⁻²   1 674973

675246

675247

676628

676632

675347

675344

676633

676634

676635

123127 Adriamycin

[0090] The present invention further provides pharmaceuticalformulations comprising an effective amount of a selenophene compoundfor treating a patient having a tumor. As used herein, an effectiveamount of the selenophene compound is defined as the amount of thecompound which, upon administration to a patient, inhibits growth oftumor cells, kills malignant cells, reduces the volume or size of thetumors or eliminates the tumor entirely in the treated patient.

[0091] The effective amount to be administered to a patient is typicallybased on body surface area, patient weight, and patient condition. Theinterrelationship of dosages for animals and humans (based on milligramsper meter squared of body surface) is described by Freireich, E. J., etal., Cancer Chemother. Rep., 50 (4): 219 (1966). Body surface area maybe approximately determined from patient height and weight (see e.g.,Scientific Tables, Geigy Pharmaceuticals, Ardley, N.Y., pages 537-538(1970)). An effective amount of the selenophene compounds in the presentinvention can range from about 5 mg/kg to about 100 mg/kg, morepreferably from about 0.25 mg/kg to about 50 mg/kg, and most preferablyabout 0.1 to about 10 mg/kg.

[0092] Effective doses will also vary, as recognized by those skilled inthe art, dependant on route of administration, excipient usage and thepossibility of co-usage with other therapeutic treatments includingother anti-tumor agents, and radiation therapy.

[0093] The pharmaceutical formulation may be administered via theparenteral route, including subcutaneously, intraperitoneally,intramuscularly and intravenously. Examples of parenteral dosage formsinclude aqueous solutions of the active agent, in a isotonic saline, 5%glucose or other well-known pharmaceutically acceptable liquid carrier.In one preferred aspect of the present embodiment, the selenophenecompound is dissolved in a saline solution containing 5% of dimethylsulfoxide and 10% Cremphor EL (Sigma Chemical Company). Additionalsolubilizing agents such as cyclodextrins, which form specific, moresoluble complexes with the present selenophene compounds, or othersolubilizing agents well-known to those familiar with the art, can beutilized as pharmaceutical excipients for delivery of the selenophenecompounds.

[0094] The present compound can also be formulated into dosage forms forother routes of administration utilizing well-known methods. Thepharmaceutical compositions can be formulated, for example, in dosageforms for oral administration in a capsule, a gel seal or a tablet.Capsules may comprise any well-known pharmaceutically acceptablematerial such as gelatin or cellulose derivatives. Tablets may beformulated in accordance with conventional procedure by compressingmixtures of the active polythiophene and solid carriers, and lubricantswell-known to those familiar with the art. Examples of solid carriersinclude starch, sugar, bentonite. The compounds of the present inventioncan also be administered in a form of a hard shell tablet or capsulecontaining, for example, lactose or mannitol as a binder andconventional fillers and tableting agents.

[0095] The following examples are provided to illustrate variousembodiments of Applicants' invention, and are not intended to in any waylimit the scope of the invention as set forth in this specification andappended claims.

EXAMPLE 1

[0096] Synthesis of α-Terselenophenes

[0097] A two-step total synthesis of α-terselenophene from selenophene(Aldrich Chemical Co.) has been developed (See Scheme 1).

[0098] Bis(tricyclohexyltin)selenide can be prepared fromtricyclohexyltin chloride (Aldrich Chemical Co.) and sodium selenide(Alfa Chemical Co.). The functional group can be introduced throughselective formylation using lithium diisopropylamide (LDA) anddimethylformamide (DMF), which can then be sequentially converted intohydroxylmethyl and aminomethyl functional groups. These functionalgroups can provide required starting points for further chemicalmodifications, see Scheme 2 as follows:

EXAMPLE 2

[0099] Synthesis of Hybrid α-Terselenophenes

[0100] The synthetic strategy designed for the preparation ofα-terselenophene can be readily modified for the synthesis of numerous“hybrid” α-selenophenes containing other five-membered heterocycles (SeeScheme 3).

[0101] Wherein X, and Y are selected from the group consisting of Se, O,S, NCH₃ and NH₂, and Z is selected from the group consisting of Se, S,NCH₃ and NH₂. Various functional groups can be introduced using theapproaches outlined in the synthesis of ∝-terselenophenes (Scheme 2).

EXAMPLE 3

[0102] Preparation of 1,4-diselenophene-1,4-diketone.

[0103] A CH₂Cl₂ solution containing selenophene (5 g) and succinylchloride (2 g) was added dropwise to an anhydrous CH₂Cl₂ solution (60mL) containing AlCl₃ (5 g) under N₂ at 0° C. The reaction mixture wasstirred at 0° C. for 1 h, slowly warmed to room temperature, and stirredfor 4 h at room temperature. The reaction mixture was poured into abeaker containing ice. Ethyl acetate (200 mL) was added and the organiclayer was separated out using a separatory funnel. The aqueous layer wasback washed with ethyl acetate (2×100 mL). The combined organic layerwas washed with H₂O (2×300 mL). The organic layer was collected, driedover MgSO₄, and the solvent was removed under vacuum. The residue waschromatographed over silica gel using (10:1) hexanes/ethyl acetate toafford the product in 25% yield.

EXAMPLE 4

[0104] Preparation of 2,2′:5′,2″-terselenophene.

[0105] A BCl₃ solution (1;0 M solution in hexanes, 580 mL) was addeddropwise to an anhydrous toluene solution (5 mL) containing1,4-diselenophene-1,4-diketone (100 mg) andbis(tricyclohexyltin)selenide (520 mg) under N₂ at room temperature. Thesolution was refluxed for 30 min and cooled to room temperature. Thereaction solution was diluted with ethyl acetate (100 mL) and washedwith H₂O (2×100 mL). The organic layer was separated, dried over MgSO₄,filtered, and the solvent was removed under vacuum. The residue waschromatographed over silica gel using hexanes to afford2,2′:5′,2″-terselenophene in 80% yield.

EXAMPLE 5

[0106] Preparation of 2-formyl-5,2′:5″,2″-terselenophene.

[0107] LDA (1.0 M solution in THF, 310 mL) was added to an anhydrous THFsolution (4 mL) containing 2,2′:5′,2″-terselenophene (100 mg) under N₂at −78° C. The solution was stirred at −78° C. for 3 h, anhydrous DMF (1mL) was added, stirred at −78° C. for-1 h, and slowly warmed to roomtemperature. The reaction solution was diluted with ethyl acetate (100mL) and washed with H₂O (4×100 mL). The organic layer was separated,dried over MgSO₄, filtered, and the solvent was removed under vacuum.The residue was chromatographed over silica gel using CH₂Cl₂ to afford2-formyl-5,2′:5′,2″-terselenophene in 75% yield.

EXAMPLE 6

[0108] Preparation of 2,5″-diformyl-5,2′:5′,2″-terselenophene.

[0109] LDA (1.0 M solution in THE, 1.0 mL) was added to an anhydrous THFsolution (4 mL) containing 2,2′:5′,2″-terselenophene (100 mg) under N₂at −78° C. The solution was stirred at −78° C. for 3 h, anhydrous DMF (2mL) was added, stirred at −78° C. for 1 h, and slowly warmed to roomtemperature. The reaction solution was diluted with ethyl acetate (100mL) and washed with H₂O (4×100 mL). The organic layer was separated,dried over MgSO₄, filtered, and the solvent was removed under vacuum.The residue was chromatographed over silica gel using (5:1) CH₂Cl₂/ethylacetate to afford 2,5″-diformyl-5,2′:5′,2″-terselenophene in 75% yield.

EXAMPLE 7

[0110] Preparation of 2-hydroxymethyl-5,2′:5′,2″-terselenophene.

[0111] NaBH₄ (10 mg) was added to a THF solution (2 mL)2-formyl-5,2′:5′,2″-terselenophene (15 mg) and stirred at roomtemperature for 2 h. The reaction solution was diluted with ethylacetate (50 mL), washed with 2N HCl (5 mL), and then washed with H₂O(3×50 mL). The organic layer was separated, dried over MgSO₄, filtered,and the solvent was removed under vacuum. The residue waschromatographed over silica gel using (2:1) hexanes/ethyl acetate toafford 2-hydroxymethyl-5,2′:5′,2″-terselenophene in 98% yield.

EXAMPLE 8

[0112] Preparation of 2,5″-dihydroxymethyl-5,2′:5′,2″-terselenophene.

[0113] NaBH₄ (10 mg) was added to a THF solution (2 mL) containing2,5″-diformyl-5,2′:5′,2″-terselenophene (15 mg) and stirred at roomtemperature for 5 h. The reaction solution was diluted with ethylacetate (50 mL), washed with 2N HCl (5 mL), and washed with H₂Q (3×100mL). The organic layer was separated, dried over MgSO₄, filtered, andthe solvent was removed under vacuum. The residue was chromatographedover silica gel using (1:1) hexanes/ethyl acetate to afford2,5″-dihydroxymethyl-5,2′:5′,2″-terselenophene in 98% yield.

EXAMPLE 9

[0114] Preparation of 2,4-diselenophenylfuran.

[0115] d-10-camphorsulfonic acid (2 g) was added to an ethanolicsolution (15 mL) containing 2′,2″-diselenophene-1,4-diketone (100 mg)and refluxed for 2 days. The reaction solution was diluted with ethylacetate (100 mL) and washed with H₂O (3×100 mL). The organic layer wasseparated, dried over MgSO₄, filtered, and the solvent was removed undervacuum. The residue was chromatographed over silica gel using (10:1)hexanes/ethyl acetate to afford 2,2′:5,2″-diselenophenylfuran in 90%yield.

EXAMPLE 10

[0116] Preparation of 5′-formyl-2,2′:5,2″-diselenophenylfuran.

[0117] LDA (1 molar solution in THF, 00 mL) was added to an anhydrousTHF solution (00 mL) containing 2,2′:5,2″-diselenophenylfuran (00 mg)under N₂ at −78° C. The solution was stirred at −78° C. for 3 h,anhydrous DMF (excess) was added, stirred at −78° C. for 1 h, and slowlywarmed to room temperature. The reaction solution was diluted with ethylacetate (00 mL) and washed with H₂O (3×100 mL). The organic layer wasseparated, dried over MgSO₄, filtered, and the solvent was removed undervacuum. The residue was chromatographed over silica gel using (4:1)hexanes/ethyl acetate to afford 5′-formyl-2,2′:5,2″-diselenophenylfuranin 00% yield.

EXAMPLE 11

[0118] Preparation of 5′,5″-diformyl-2,2′:5,2″-diselenophenylfuran.

[0119] LDA (1 molar solution in THF, 00 mL) was added to an anhydrousTHF solution (00 mL) containing 2,2′:5,2″-diselenophenylfuran (00 mg)under N₂ at −78° C. The solution was stirred at −78° C. for 3 h,anhydrous DMF (excess) was added, stirred at −78° C. for 1 h, and slowlywarmed to room temperature: The reaction solution was diluted with ethylacetate (00 μL) and washed with H₂O (3×100 mL). The organic layer wasseparated, dried over MgSO₄, filtered, and the solvent was removed undervacuum. The residue was chromatographed over silica gel using (4:1)hexanes/ethyl acetate to afford5,5″-diformyl-2,2′:5,2″-diselenophenylfuran in 00% yield.

EXAMPLE 12

[0120] Preparation of 5′-hydroxymethyl-2,2′:5,2″-diselenophenylfuran.

[0121] NaBH₄ (excess) was added to a THF solution (00 mL) containing5′-formyl-2,2′:5,2″-diselenophenylfuran (00 mg) and stirred at roomtemperature for 5 h. The reaction solution was diluted with ethylacetate (100 mL) and washed with H₂O (3×100 mL). The organic layer wasseparated, dried over MgSO₄, filtered, and the solvent was removed undervacuum. The residue was chromatographed over silica gel using (2:1)hexanes/ethyl acetate to afford5′-hydroxymethyl-2,2′:5,2″-diselenophenylfuran in 00% yield.

EXAMPLE 13

[0122] Preparation of5′,5″-dihydroxymethyl-2,2′:5,2″-diselenophenylfuran.

[0123] NaBH₄ (excess) was added to a THF solution (00 mL) containing5′,5″-diformyl-2,2′:5,2″-diselenophenylfuran (00 mg) and stirred at roomtemperature for 5 h. The reaction solution was diluted with ethylacetate (100 mL) and washed with H₂O (3×100 mL). The organic layer wasseparated, dried over MgSO₄, filtered, and the solvent was removed undervacuum. The residue was chromatographed over silica gel using (1:1)hexanes/ethyl acetate to afford5′,5″-dihydroxymethyl-2,2′:5,2″-diselenophenylfuran in 00% yield.

EXAMPLE 14

[0124] Preparation of 2,1′:5,2″-diselenophenylthiophene.

[0125] BCl₃ (1.0 M solution in hexanes, 580 mL) was added dropwise to ananhydrous toluene solution (5 mL) containing2′,2″-diselenophenyl-1,4-diketone (100 mg) andbis(tricyclohexyltin)sulfide (520 mg) under N₂ at room temperature. Thesolution was refluxed for 30 min and cooled to room temperature. Thereaction solution was diluted with ethyl acetate (100 mL) and washedwith H₂O (2×100 mL). The organic layer was separated, dried over MgSO₄,filtered, and the solvent was removed under vacuum. The residue waschromatographed over silica gel using hexanes to afford2,2′:5,2″-diselenophenylthiophene in 85% yield.

EXAMPLE 15

[0126] Preparation of 5′-formyl-2,2′:5,2″-diselenophenylthiophene.

[0127] LDA (1.0 M solution in THF, 350 mL) was added to an anhydrous THFsolution (4 mL) containing 2,2′:5,2″-diselenophenylthiophene under N₂ at−78° C. The solution was stirred at −78° C. for 3 h, anhydrous DMF (1mL) was added, stirred at −78° C. for 1 h, and slowly warmed to roomtemperature. The reaction solution was diluted with ethyl acetate (50mL) and washed with H₂O (3×100 mL). The organic layer was separated,dried over MgSO₄, filtered, and the solvent was removed under vacuum.The residue was chromatographed over silica gel using CH₂Cl₂ to afford5′-formyl-2,2′:5,2″-diselenophenylthiophene in 80% yield.

EXAMPLE 16

[0128] Preparation of 5′,5″-diformyl-2,2′:5,2″-diselenophenylthiophene.

[0129] LDA (1.0 M solution in THF, 1 mL) was added to an anhydrous THFsolution (4 mL) containing 2,2′:5,2″-diselenophenylthiophene under N₂ at−78° C. The solution was stirred at −78° C. for 3 h, anhydrous DMF (2mL) was added, the solution was stirred at −78° C. for 1 h, and slowlywarmed to room temperature. The reaction solution was diluted with ethylacetate (100 mL) and washed with H₂O (3×100 mL). The organic layer wasseparated, dried over MgSO₄, filtered, and the solvent was removed undervacuum. The residue was chromatographed over silica gel using (5:1)CH₂Cl₂/ethyl acetate to afford5′,5″-diformyl-2,2′:5,2″-diselenophenylthiophene in 80% yield.

EXAMPLE 17

[0130] Preparation of5′-hydroxymethyl-2,2′:5,2″-diselenophenylthiophene.

[0131] NaBH₄ (10 mg) was added to a THF solution (3 mL) containing5′-formyl-2,2′:5,2″-diselenophenylthiophene (20 mg) and stirred at roomtemperature for 2 h. The reaction solution was diluted with ethylacetate (50 mL) and washed with H₂O (5×50 mL). The organic layer wasseparated, dried over MgSO₄, filtered, and the solvent was removed undervacuum. The residue was chromatographed over silica gel using (2:1)CH₂Cl₂/ethyl acetate to afford5′-hydroxymethyl-2,2′:5,2″-diselenophenylthiophene in 98% yield.

EXAMPLE 18

[0132] Preparation of55″-dihydroxymethyl-2,2′:5,2″-diselenophenylthiophene.

[0133] NaBH₄ (10 mg) was added to a THF solution (3 mL) containing5′,5″-diformyl-2,2′:5,2″-diselenophenylthiophene (20 mg) and stirred atroom temperature for 5 h. The reaction solution was diluted with ethylacetate (50 mL) and washed with H₂O (5×50 mL). The organic layer wasseparated, dried over MgSO₄, filtered, and the solvent was removed undervacuum. The residue was chromatographed over silica gel using (1:1)hexanes/ethyl acetate to afford5′,5″-dihydroxymethyl-2,2′:5,2″-diselenophenylthiophene in 98% yield.

EXAMPLE 19

[0134] Preparation of 2,2′:5,2″-diselenophenylpyrrole.

[0135] An ethanolic solution (20 mL) containing2′,2″-diselenophenyl-1,4-diketone (200 mg) and ammonium acetate (500 mg)and sodium acetate (200 mg) was refluxed overnight. The reactionsolution was diluted with ethyl acetate (100 mL) and washed with H₂O(3×100 mL). The organic layer was separated, dried over MgSO₄, filtered,and the solvent was removed under vacuum. The residue waschromatographed over silica gel using (10:1) hexanes/ethyl acetate toafford 2,2′:5,2″-diselenophenylpyrrole in 94% yield.

EXAMPLE 20

[0136] Preparation of 5′-formyl-2,2′:5,2″-diselenophenylpyrrole.

[0137] LDA (1.0 M solution in THF, 760 mL) was added to an anhydrous THFsolution (5 mL) containing 2,2′:5,2″-diselenophenylpyrrole (100 mg)under N₂ at −78° C. The solution was stirred at −78° C. for 3 h,anhydrous DMF (1.5 mL) was added, the solution was slowly warmed to roomtemperature, and stirred at room temperature for 2 h. The reactionsolution was diluted with ethyl acetate (100 mL) and washed with H₂O(3×100 mL). The organic layer was separated, dried over MgSO₄, filtered,and the solvent was removed under vacuum. The residue waschromatographed over silica gel using (3:1) hexanes/ethyl acetate toafford 5′-formyl-2,2′:5,2″-diselenophenylpyrrole in 75% yield.

EXAMPLE 21

[0138] Preparation of 5′-hydroxymethyl-2,2′:5,2″-diselenophenylpyrrole.

[0139] NaBH₄ (20 mg) was added to a THE solution (2 mL) containing5′-formyl-2,2′:5,2″-diselenophenylpyrrole (20 mg) and stirred at roomtemperature for 2 h. The reaction solution was diluted with ethylacetate (50 mL) and washed with H₂O (3×50 mL). The organic layer wasseparated, dried over MgSO₄, filtered, and the solvent was removed undervacuum. The residue was chromatographed over silica gel using (2:1)hexanes/ethyl acetate to afford5′-hydroxymethyl-2,2′:5,2″-diselenophenylpyrrole in 98% yield.

EXAMPLE 22

[0140] Preparation of 2′,2″-difuranyl-1,4-diketone.

[0141] A CH₂Cl₂ solution containing furan (10 mL) and succinyl chloride(2 g) was added dropwise to an anhydrous CH₂Cl₂ solution (100 mL)containing AlCl₃ (10 g) under N₂ at 0° C. The reaction mixture wasstirred at 0° C. for 2 h, slowly warmed to room temperature, and stirredfor 4 h. The reaction mixture was poured into a beaker containing ice.Ethyl acetate (300 mL) was added and the organic layer was separated outusing a separatory funnel. The aqueous layer was back washed with ethylacetate (2×100 mL). The combined organic layer was washed with H₂O(2×300 mL). The organic layer was collected, dried over MgSO₄, and thesolvent was removed under vacuum. The residue was chromatographed oversilica gel using (3:1) hexanes/ethyl acetate to afford2′,2″-difuranyl-1,4-diketone in 25% yield.

EXAMPLE 23

[0142] Preparation of 2,2′:5,2″-difuranylselenophene.

[0143] BCl₃ (1.0 M solution in hexanes, 900 mm) was added dropwise to ananhydrous, toluene solution (00 mL) containing2′,2″-difuranyl-1,4-diketone (100 mg) and bis(tricyclohexyltin)-selenide(750 mg) under N₂ at room temperature. The solution was refluxed for 30min and cooled to room temperature. The reaction solution was dilutedwith ethyl acetate (100 mL) and washed with H₂O (2×100 mL). The organiclayer was separated, dried over MgSO₄, filtered, and the solvent wasremoved under vacuum. The residue was chromatographed over silica gelusing hexanes to afford 2,2′:5,2″-difuranylselenophene in 80% yield.

EXAMPLE 24

[0144] Preparation of 5′-formyl-2,2′:5,2″-difuranylselenophene.

[0145] LDA (1.0 M solution in THF, 420 mL) was added to an anhydrous THFsolution (4 mL) containing 2,2′:5,2″-difuranylselenophene (100 mg) underN₂ at −78° C. The solution was stirred at −78° C. for 3 h, anhydrous DMF(1 mL) was added, the solution was stirred at −78° C. for 1 h, andslowly warmed to room temperature. The reaction solution was dilutedwith ethyl acetate (100 mL) and washed with H₂O (3×100 mL). The organiclayer was separated, dried over MgSO₄, filtered, and the solvent wasremoved under vacuum. The residue was chromatographed over silica gelusing (3:1) hexanes/ethyl acetate to afford5′-formyl-2,2′:5,2″-difuranylselenophene in 75% yield.

EXAMPLE 25

[0146] Preparation of 5′,5″-diformyl-2,2′:5,2″-difuranylselenophene.

[0147] LDA (1.0 M solution in TH, 00 mL) was added to an anhydrous THsolution (4 mL) containing 2,2′:5,2″-difuranylselenophene (100 mg) underN₂ at −78° C. The solution was stirred at −78° C. for 3 h, addedanhydrous DMF (2 mL), stirred at −78° C. for 1 h, and slowly warmed toroom temperature. The reaction solution was diluted with ethyl acetate(100 mid) and washed with H₂O (3×100 nm). The organic layer wasseparated, dried over MgSO₄, filtered, and the solvent was removed undervacuum. The residue was chromatographed over silica gel using (2:1)hexanes/ethyl acetate to afford5′,5″-diformyl-2,2′:5,2″-difuranylselenophene in 80% yield.

EXAMPLE 26

[0148] Preparation of 5′-hydroxymethyl-2,2′:5,2″-difuranylselenophene.

[0149] NaBH₄ (10 mg) was added to a THF solution (2 mL) containing5′-formyl-2,2′:5,2″-difuranylselenophene (20 mg) and stirred at roomtemperature for 5 h. The reaction solution was diluted with ethylacetate (50 mL) and washed with H₂O (3×50 mL). The organic layer wasseparated, dried over MgSO₄, filtered, and the solvent was removed undervacuum. The residue was chromatographed over silica gel using (2:1)hexanes/ethyl acetate to afford5′-hydroxymethyl-2,2′:5,2″-difuranylselenophene in 98% yield.

EXAMPLE 27

[0150] Preparation of5′,5″-dihydroxymethyl-2,2′:5,2″-difuranylselenophene.

[0151] NaBH₄ (10 mg) was added to a THF solution (2 mL) containing5′,5″-diformyl-2,2′:5,2″-difuranylselenophene (20 mg) and stirred atroom temperature for 5 h. The reaction solution was diluted with ethylacetate (50 mL) and washed with H₂O (3×50 mL). The organic layer wasseparated, dried over MgSO₄, filtered, and the solvent was removed undervacuum. The residue was chromatographed over silica gel using (1:1)hexanes/ethyl acetate to afford5′,5″-dihydroxymethyl-2,2′:5,2″-difuranylselenophene in 98% yield.

EXAMPLE 28

[0152] Preparation of 2′,2″-dithienyl-1,4-diketone.

[0153] A CH₂Cl₂ solution-containing thiophene (10 mL) and succinylchloride (2 g) was added dropwisely to an anhydrous CH₂Cl₂ solution (100mL) containing AlCl₃ (10 g) under N₂ at 0° C. The reaction mixture wasstirred at 0° C. for 2 h, slowly warmed to room temperature, and stirredfor 4 h. The reaction mixture was poured into a beaker containing ice.Ethyl acetate (300 mL) was added and the organic layer was separated outusing a separatory funnel. The aqueous layer was back washed with ethylacetate (2×100 mL). The combined organic layer was washed with H₂O(2×300 mL). The organic layer was collected, dried over MgSO₄, and thesolvent was removed under vacuum. The residue was chromatographed oversilica gel using (3:1) hexanes/ethyl acetate to afford2′,2″-dithienyl-1,4-diketone in 25% yield.

EXAMPLE 29

[0154] Preparation of 2,2′:5,2″-dithienylselenophene.

[0155] BCl₃ (1.0 M solution in hexanes, 1.6 mL) was added dropwise to ananhydrous toluene solution (5 mL) containing2′,2″-dithienyl-1,4-diketone (200 mg) and bis(tricyclohexyltin)selenide(1.3 g) under N₂ at room temperature. The solution was refluxed for 30min and cooled to room temperature. The reaction solution was dilutedwith ethyl acetate (100 mL) and washed with H₂O (3×100 mL). The organiclayer was separated, dried over MgSO₄, filtered, and the solvent wasremoved under vacuum. The residue was chromatographed over silica gelusing hexanes to afford. 2,2′:5,2″-dithienylselenophene in 90% yield.

EXAMPLE 30

[0156] Preparation of 5′-formyl-2,2′:5,2″-dithienylselenophene.

[0157] LDA (1.0 M solution in THF, 380 mL) was added to an anhydrousTIFF solution (4 mL) containing 2,2′:5,2″-dithienylselenophene (100 mg)under N₂ at −78° C. The solution was stirred at −78° C. for 3 h,anhydrous DMF (1 mL) was added, the solution stirred at −78° C. for 1 h,and slowly warmed to room temperature. The reaction solution was dilutedwith ethyl acetate (100 mL) and washed with H₂O (3×100 mL). The organiclayer was separated, dried over MgSO₄, filtered, and the solvent wasremoved under vacuum. The residue was chromatographed over silica gelusing (3:1) hexanes/ethyl acetate to afford5′-formyl-2,2′:5,2″-dithienylselenophene in 75% yield.

EXAMPLE 31

[0158] Preparation of 5′,5″-diformyl-2,2′:5,2″-dithienylselenophene.

[0159] LDA (1.0 M solution in THF, 1.0 mL) was added to an anhydrous THFsolution (4 mL) containing 2,2′:5,2″-dithienylselenophene (100 mg) underN₂ at −78° C. The solution was stirred at −78° C. for 3 h, anhydrous DMF(2 mL) was added, the solution stirred at −78° C. for 1 h, and slowlywarmed to room temperature. The reaction solution was diluted with ethylacetate (100 mL) and washed with H₂O (3×100 mL). The organic layer wasseparated, dried over MgSO₄, filtered, and the solvent was removed undervacuum. The residue was chromatographed over silica gel using (2:1)hexanes/ethyl acetate to afford5′,5″-diformyl-2,2′:5,2″-dithienylselenophene in 85% yield.

EXAMPLE 32

[0160] Preparation of 5′-hydroxymethyl-2,2′:5,2″-dithienylselenophene.

[0161] NaBH₄ (10 mg) was added to a THF solution (2 mL) containing5′-formyl-2,2′:5,2″-dithienylselenophene (20 mg) and stirred at roomtemperature for 5 h. The reaction solution was diluted with ethylacetate (50 mL) and washed with H₂O (3×50 mL). The organic layer wasseparated, dried over MgSO₄, filtered, and the solvent was removed undervacuum. The residue was chromatographed over silica gel using (2:1)hexanes/ethyl acetate to afford5′-hydroxymethyl-2,2′:5,2″-dithienylselenophene in 98% yield.

EXAMPLE 33

[0162] Preparation of5′,5″-dihydroxymethyl-2,2′:5,2″-dithienylselenophene.

[0163] NaBH₄ (10 mg) was added to a THF solution (2 mL) containing5′,5″-diformyl-2,2′:5,2″-dithienylselenophene (20 mg) and stirred atroom temperature for 5 h. The reaction solution was diluted with ethylacetate (50 mL) and washed with H₂O (3×50 mL). The organic layer wasseparated, dried over MgSO₄, filtered, and the solvent was removed undervacuum. The residue was chromatographed over silica gel using (1:1)hexanes/ethyl acetate to afford5′,5″-dihydroxymethyl-2,2′:5,2″-dithienylselenophene in 98% yield.

EXAMPLE 34 Alternative Method of Synthesizing Hybrid α-Terselenophenes

[0164] In addition to the method of synthesis described in Example 2, analternative synthesis strategy (Scheme 4) can be utilized to preparenumerous “hybrid” α-Terselenophenes.

[0165] Wherein X, and Y are selected from the group consisting of Se, O,S, N(CH₃) and NH, and Z is selected from the group consisting of Se, S,N(CH₃) and NH. Various functional groups can be introduced using theapproaches outlined in the synthesis of terselenophenes (Scheme 2).

EXAMPLE 35 Preparation of 3-Dimethylamino-1-(2′-selenyl)propanone

[0166]

[0167] Synthesis of 2-acetylselenophene (1): A solution of selenophene(2.0 g, 15 mmol), acetic anhydride (2.34 g, 23 mmol) and tin (IV)chloride (0.06 g, 0.23 mmol) in 30 mL of dry methene chloride wasstirred under Argon for two days until TLC plate showed completion ofthe reaction.

[0168] A mixture of crude 2-acetylselenophene (2.6 g, 15 mmol),paraformaldehyde (0.59 g, 19.6 mmol), dimethylamine hydrochloride (1.6g, 19.5 mmol) and 0.15 mL of HCl was refluxed for 16 h in 7 mL ofethanol. The reaction mixture was cooled and the precipitate wasfiltered, washed with ether and dried; yield 2.77 g (69.3%). ThisMannich base hydrochloride (2 g) was basified using ammonium hydroxide.The solution was extracted (3×15 mL) with diethyl ether. The organiclayer was washed with water and dried with sodium sulfate. Evaporationof ether gave 1.6 g of product 2.

[0169]¹H NMR (CDCl₃) δ 8.37 (dd, 1H, H-5 of selenophene ring, J=5.52,0.99), 7.95 (dd, 1H, H-3 of selenophene ring, J=0.99, 3.99), 7.40 (dd,1H, H-4 of selenophene ring, J=5.52, 3.99), 3.10 (t, 2H, CO—CH₂, J=7.6),2.76 (t, 2H, CH₂—NMe₂, J=7.6), 2.29 (s, 6H, NMe₂).

EXAMPLE 36 Preparation of1-(2′-Thienyl)-4-(2″-selenyl)butane-1,4-dione(3)

[0170]

[0171] A solution of 2-formylthiophene (1.05 g, 9.4 mmol) in 4 mL dryDMF was added to a suspension of sodium cyanide (0.16 g, 3.4 mmol) in 4mL, dry DMF After stirring for 10 min, the3-dimethylamino-1-(2′-selenyl)propanone 2 (1.73 g, 7.52 mmol) in 10 mLDMF was added slowly. The mixture was stirred overnight. Water was added(30 ML), and the product was extracted with chloroform (3×30 mL). Theextract was washed with water, dried over sodium sulfate, andevaporated. The product 3 was recrystallized from ethanol; yield: 1.97 g(88.3%). mp: 121-122.40° C. ¹H NMR (500 MHz, CDCl₃) δ 8.37 (dd, 1H, H-5of selenophene ring, J=5.46, 0.91), 8.03 (dd, 1H, H-3 of selenophenering, J=3.92, 0.91), 7.81 (dd, 1H, H-5 of thiophene ring, J=0.94, 3.82),7.63 (dd, 1H, H-3 of thiophene ring, J=4.91,0.94), 7.40 (dd, 1H, H-4selenophene ring, J=5.46, 0.91), 7.14 (dd, 1H, H-4 of thiophene ring,J=3.82, 4.91), 3.40 (m, 4H, CH₂—CH₂). Anal. Calcd. for C₁₂H₁₀O₂SSe: C,48.49; H, 3.39; S, 10.79. Found: C, 48.83; H, 3.38; S, 10.46.

EXAMPLE 37 Preparation of 2-(2′-Selenyl)-5-(2″-thienyl)thiophene (4)

[0172]

[0173] 1-(2′-Thienyl)-4-(2″-selenyl)butane-1,4-dione 3 (1.1 g, 3.70mmol) and Lawesson's reagent (0.99 g, 2.44 mmol) were refluxed overnightin 15 mL toluene. The toluene was evaporated and the crude product waspurified using silica flash column with ether/hexane as eluent. Theproduct 4 was recrystallized from methanol; yield: 0.9 g (82.3%). mp.103-104° C. ¹H NMR (CDCl₃) δ 7.84 (dd, 1H, J=5.57, 1.04), 7.30 (dd, 1H,J=3.78,1.04), 7.20 (m, 2H), 7.15 (dd, 1H, J=3.52, 1.1), 7.00 (m, 3H);¹³C NMR (300 Mhz, CDCl₃) δ 142.09 (weak), 138.36 (weak), 137.01 (weak),136.32 (weak), 130.29, 129.60, 127.84, 125.73, 124.96, 124.45, 124.29,123.63. Anal. Calcd for C₁₂H₈S₂Se: C, 48.81; H, 2.73; S, 21.72. Found:C, 49.19; H, 2.58; S, 21.68.

EXAMPLE 38 Preparation of 2-(2′-Selenyl)-5-(2″-thienyl)furene (5)

[0174]

[0175] 1-(2′-Thienyl)-4-(2″-selenyl)butane-1,4-dione 3 (0.76 g, 2.56mmol) was added to 35 mL of acetic anhydride, then slowly added 3.0 mlof HCl After 4 h at room temperature, the reaction mixture was pouredinto ice water and extracted with ether. The organic layer was washedwith NaHCO₃ and water, dried over sodium sulfate. After evaporation ofthe solvent, the crude material was subjected to silica columnpurification to give the product 5. Yield: 0.51 g (75.5%). The yellowishwhite solid was recrystallized from methanol. mp 85-87° C. ¹HNMR (CDCl₃)δ 7.89 (dd, 1H, J=4.51, 1.03), 7.44 (dd, 1H, J=3.82, 1.01), 7.29(dd, 1H,J=3.72, 1.08), 7.26 (dd, 1H, J=4.51, 3.82), 7.22 (dd, 1H, J=5.03, 1.08),7.03 (dd, 1H, J=5.02, 3.70), 6.53 (m, 2H).

EXAMPLE 39 Preparation of 2-(2′-Selenyl)-5-(2″-thienyl)pyrrol (6)

[0176]

[0177] 1-(2′-Thienyl)-4-(2″-selenyl)butane-1,4-dione 3 (0.4 g, 1.35mmol), sodium acetate (0.33 g, 4.0 mmol) and ammonium acetate (0.78 g,10.1 mmol) were refluxed at 95° C. overnight in 20 mL ethanol. Thesolvent was evaporated and the crude product 6 was purified using silicaflash column with ether/hexane as eluent; yield: 0.27 g (73%). mp.82-83.5° C. ¹H NMR δ 8.26 (br, 1H), 7.81 (d, 1H, J=5.27), 7.25 (dd, 1H,J=3.78, 5.27), 7.20(d, 1H, J=3.78), 7.16 (d, 1H, J=5.01), 7.07 (d, 1H,J=3.60), 7.02 (dd, 1H, J=5.01, 3.60), 6.40 (m, 2H).

EXAMPLE 40 Preparation of 1-(2′-Selenyl)-4-(2″-furyl)butane-1,4-dione(7)

[0178]

[0179] A solution of 2-formylfurene (2.27 g, 23.65 mmol) in 20 mL dryDMF was added to a suspension of sodium cyanide (0.42 g, 8.45 mmol) in10 mL dry DMF After stirring for 10 min,3-dimethylamino-1-(2′-selenyl)propanone 2 (4.3 g, 18.8 mmol) in 20 mLDMF was added slowly. The mixture was stirred overnight. Water was added(100 mL), and the product was extracted with chloroform (3×100 mL). Theextract was washed with water, dried over sodium sulfate, andevaporated. The product 7 was recrystallized from ethanol; yield: 3.52 g(66.7%). mp. 82-83.5° C. ¹H NMR (CDCl₃) δ 8.35 (dd, 1H, H-5 ofselenophene ring, J=5.51, 0.78), 8.01 (dd, 1H, H-3 of selenophene ring,J=3.99, 0.79), 7.58 (d, 1H, H-5 of furene ring, J=1.71), 7.39 (dd, 1H,H-4 of selenophene ring, J=5.52, 3.99), 7.23 (d, 1H, H-3 of furene ring,J=3.54), 6.53 (dd, 1H, H-4 of furene ring, J=3.54, 1.70), 3.33 (m, 4H,CH₂—CH₂).

EXAMPLE 41 Preparation of 2-(2′-Selenyl)-5-(2″-furyl)thiophene (8)

[0180]

[0181] 1-(2′-Selenyl)-4-(2″-furyl)butane-1,4-dione 7 (0.25 g, 0.9 mmol)and Lawesson's reagent (0.66 g, 1.63 mmol) were refluxed overnight in 7mL toluene. The toluene was evaporated and the crude product waspurified using silica flash column with ether/hexane as eluent. Theproduct 8 was recrystallized from methanol; yield: 0.22 g (88%). mp.76-77° C. ¹H NMR δ 7.86 (dd, 1H, J=5.58, 1.00), 7.40 (d, 1H, J=1.76),7.31(dd, 1H, J=3.87, 1.00), 7.23 (dd, 1H, J=5.59, 3.87), 7.11 (d, 1H,J=3.78), 7.03 (d, 1H, J=3.81), 6.49 (d, 1H, J=3.36), 6.43(dd, 1H,J=1.77, 3.36).

EXAMPLE 42 Preparation of 2-(2′-Selenyl)-5-(2″-furyl)pyrrol (9)

[0182]

[0183] 1-(2′-Thienyl)-4-(2″-furyl)butane-1,4-dione 7 (0.20 g, 0.71mmol), sodium acetate (0.18 g, 2.1 mmol) and ammonium acetate (0.41 g,5.3 mmol) were refluxed at 95° C. overnight in 12 mL ethanol. Thesolvent was evaporated and the crude product 9 was purified using silicaflash column with ether/hexane as eluent; yield: 0.15 g (80%). mp.73-74° C. ¹HNMR δ 8.50 (br, 1H), 7.80 (d, 1H, J=5.45), 7.36 (dd, 1H,J=1.02, 0.78), 7.22(m, 2H), 6.40 (in, 4H).

EXAMPLE 43 Preparation of 1,4-Bis-(2′-selenyl)butane-1,4-dione (11)

[0184]

[0185] Synthesis of 2-formylselenophene (10): A solution of selenophene(1.31 g, 10 mmol) in 10 ml dichloroethane was added to a mixture offreshly distilled phosphorus oxychloride (2.0 g, 13 mmol) and DMF (1.10g, 15 mmol). After stirring for 12 hr at 60° C., 2 ml water solution ofsodium acetate (2.04 g, 15 mmol) was added to the reaction mixture, andthe mixture was allowed to react for another hour. Water was added (20mL), and the product was extracted with dichloromethane (3×20 mL). Theextract was washed with water, dried over sodium sulfate, and carefullyevaporated.

[0186] A solution of crude 2-formylselenophene (454 mg, 2.9 mmol) in 0.6mL dry DMF was added to a suspension of sodium cyanide (34.3 mg, 0.7mmol) in 0.4 mL dry DMF. After stirring for 5 min, the Mannich base,3-dimethylamino-1-(2-selenyl)propanone 2 (368 mg, 1.6 mmol) in 1.2 mLDMF was added slowly. The mixture was stirred overnight. Water was added(4 mL, and the product was extracted with dichloromethane (3×6 mL). Theextract was washed with water, dried over sodium sulfate, andevaporated. The product was purified from silica gel chromatography withTHF/hexane as eluent. Yield: 105 mg (20%). ¹HNMR (CDCl₃) δ 8.37 (dd,1H×2, H-5 of selenophene ring, J=5.53, 1.01), 8.02 (dd, 1H×2, H-3 ofselenophene ring, J=1.00, 3.97), 7.40 (dd, 1H×2, H-4 of selenophenering, J=3.97, 5.50), 3.39 (s, 2H×2, CH₂—CH₂).

EXAMPLE 44 Preparation of 2,5-Bis-(2′-selenyl)-N-methylpyrrol (12)

[0187]

[0188] 1,4-Bis-(2′-selenyl)butane-1,4-dione 11 (34.4 mg, 0.1 mmol),sodium acetate (123 mg, 0.15 mmol) and methylamine chloride (101.3 mg,0.15 mmol) were refluxed overnight in 3 mL ethanol. 10 ml water was thenadded, and the product was extracted with dichloromethane. The product12 was recrystallized from ethanol; yield: 26 mg (76%). ¹H NMR (CDCl₃) δ7.96 (dd, 1H×2, H-5 of selenophene ring J=5.64, 1.64), 7.31 (dd, 1H×2,H-4 of selenophene ring J=5.64, 3.78), 7.20 (dd, 1H×2, H-3 ofselenophene ring J=3.78, 1.64), 6.32 (s, 1H×2, H-3/4 of pyrrol ring),3.74 (s, 3H, N-Me).

EXAMPLE 45 Preparation of 2,5-Bis-(2′-thienyl)selenophene (13)

[0189]

[0190] Selenophene (22 mg, 0.28 mmol) and sodium (19.2 mg, 0.83 mmol)were stirred under argon in dry DMF (10 ml at 100° C. until the solutiondecolorized, forming a brown suspension (2 h). The mixture of MeOH andEtOH (1:1, 2 mL) was added to the suspension at 0° C., followed byaddition of 1,4-Bis(2-thienyl)butadiyne (30 mg, 0.139 mmol) in solutionof THF (3 mL). After half hour, the mixture was then poured into water(20 mL) and extracted with ether (3×15 mL). The concentrated organiclayer yielded 28 mg of 13 (68.7%) after silica chromatography. ¹NMR wasidentical with the literature. (R. Shabana et al. Phosphorus, Sulfur,and Silicon, 1990, 48, 239-244).

EXAMPLE 46 Preparation of 2,5-Bis-(2′-furyl)selenophene (14)

[0191]

[0192] Selenophene (868 mg 11 mmol) and sodium (757 mg, 33 mmol) werestirred under argon in dry DMF (15 mL) at 100° C. until the solutiondecolorized, forming a brown suspension (2 h). The mixture of MeOH andEtOH (1:1, 3 mL) was added to the suspension at 0° C., followed byaddition of 1,4-Bis-(2′-furyl)butadiyne (1 g, 5.5 mmol) in solution ofTHF (3 mL). After half hour, the mixture was poured into water (20 mL)and extracted with ether (3×20 mL). The concentrated organic layeryielded 0.343 g (24%) of 14 after silica chromatography with hexane aseluent. ¹H NMR was identical with the literature.(R. Shabana et al.Phosphorus, Sulfur, and Silicon, 1990, 48, 239-244).

EXAMPLE 47 Preparation of 5′-Formyl-2,5-bis-(2′-furyl)selenophene (15)

[0193]

[0194] To a solution of 2,5-bis-(2′-furyl)selenophene 14 (0.12 g, 0.456mmol) in THF, lithium diisopropyl amide (0.73 mmol) was added at −78° C.under argon. The mixture was stirred below −20° C. for 3 h. A largeexcess of DMF (6.5 mmol) was added at −78° C., and the mixture wasallowed to gradually rise to room temperature. Ether (10 mL) was added,and the organic solution was washed with water, dried over sodiumsulfate, and evaporated. The crude solid was purified by flash columnchromatography over silica gel (ether/hexane) to give monosubstitutedaldehydes 15. Yield: 78 mg (60%), which was recrystallized fromTHF/Hexane to provide pure product. mp: 87.5-89.2 C. ¹HNMR(CDCl₃) δ 9.58(s, 1H), 7.58 (d, 1H, J=4.04), 7.43 (s, 1H), 7.35 (d, 1H, J=4.04), 7.26(d, 1H, J=3.69), 6.64 (d, 1H, J=3.69), 6.57 (d, 1H, J=3.16), 6.46 (m,1H).

EXAMPLE 48 Preparation of 5′-Hydroxymethyl-2,5-bis-(2′-furyl)selenophene(16)

[0195]

[0196] To a solution of 5′-formyl-2,5-bis-(2′-furyl)selenophene (15 mg,0.05 mmol) in 5 ml THF/MeOH (1:1), excess NaBH₄ was added at roomtemperature. The solution was stirred for 2 h. Ethyl acetate was added,and the organic solution was washed with water, dried over sodiumsulfate, and evaporated. The crude solid was purified byrecrystallization from THF/Hexane to provide pure product 16. Yield: 14mg (93.4%). mp: 75.0-77.4 C. ¹HNMR (CDCl₃) δ0.39 (in, 1H), 7.31 (in,2H), 6.48 (in, 1H), 6.43 (in, 2H), 6.33 (in, 1H). ¹³C NMR (THF-d₈) δ153.36 (weak), 150.99 (weak), 141.81, 136.66 (weak), 136.19 (weak),125.46 (weak), 124.99, 124.71, 111.99, 105.89, 105.15, 57.43.

EXAMPLE 49 Preparation of5′,5″-Diformyl-2-(2′-selenyl)-5-(2″-thienyl)thiophene (17)

[0197]

[0198] To a solution of 2-(2′-selenyl)-5-(2″-thienyl)thiophene 4 (0.45g, 1.53 mmol) in THF was added lithium diisopropyl amide (2.44 mmol) at−78° C. under argon. The mixture was stirred below −20° C. for 3 h.Large excess of DMF (13 mmol) was added at −78° C., and the mixture wasallowed to gradually rise to room temperature. Ether (30 mL) was added,and the organic solution was washed with water, dried over sodiumsulfate, and evaporated. The crude solid was purified by flash columnchromatography over silica gel (ether/hexane) to give disubstitutedaldehydes 17. Yield: 135 mg (27.3%), which was recrystallized fromTHF/Hexane to provide pure product. mp: 197.8-199.0° C. ¹H NNIR (CDCl₃)δ 9.88 (s, 1H), 9.75 (s, 1H), 7.92 (d, III, J=4.28), 7.69 (d, 1H,J=3.87), 7.46 (d, 1H, J=4.28), 7.30 (d, 1H, J=1.93), 7.29 (d, 1H,J=1.93), 7.26 (d, III, J=3.87).

EXAMPLE 50 Preparation of5′,5″-Dihydroxymethyl-2-(2′-selenyl)-5-(2″-thienyl)-thiophene (18)

[0199]

[0200] To a solution of5′,5″-diformyl-2-(2′-selenyl)-5-(2″-thienyl)thiophene (12 mg, 0.03 mmol)in 1.5 ml THF/MeOH (1:1), excessive NaBH₄ was added at room temperature.The solution was stirred for 4 h. Ethyl acetate was added, and theorganic solution was washed with water, dried over sodium sulfate, andevaporated. The crude solid was purified by recrystallization fromTHF/Hexane to provide pure product 18. Yield: 8.2 mg (68.3%). mp:187.1-188.8° C. ¹ H NNM (CDCl₃) δ 7.20 (d,IH, J=3.76), 7.07 (m, 3H),7.00 (d, 1H, J=3.66), 6.88 (d, 1H, J=3.39), 5.56 (t, 1H, OH), 5.45 (t,1H,OH), 4.65 (m, 4H, 2CH₂).

EXAMPLE 51 Synthesis of Water Soluble Analogs

[0201] A highly polar functional group can be incorporated into theselenophene compounds in order to improve their water solubility.Addition of a carbonylic functional group through an ester linkage(Scheme 5) resulted in a transient solubility. However, the benzylicester may be readily hydrolyzed to regenerate the water insolublestarting material.

[0202] On the basis of the synthesis for hybrid α-terselenophenes(Scheme 3), a nitrogen atom can be introduced into the five-memberedring system (Scheme 6). Conversion of the hydroxyl group of theintermediate compound of scheme 3 into an amino group can improve watersolubility. Further modification of its formulation may further enhancesolubility to >1 mg/ml H₂O; The ammonium analog should be highly watersoluble.

[0203] To maximize the efficiency of synthesizing hybridα-terselenophenes, Scheme 1 can be modified to produce relatedselenophene analogs in accordance with Scheme 7:

EXAMPLE 52

[0204] Synthesis of Prodrugs

[0205] An alternative approach of enhancing the water solubility ofhydrophobic drugs comprises the preparation of their polar prodruganalogs.

[0206] a. Glycosides: Preliminary results indicate that β-D-glucoside of2-hydroxymethyl-α-terthiophene retains both its in vitro and in vivoactivities. Scheme 8 illustrates a procedure utilized for the synthesisof glucoside, galactoside or, glucuronic acid analogs ofα-terselenophene:

[0207] b. Glutamate Conjugate: As mentioned above, conversion of thehydroxyl group of 2-hydroxymethyl-5,2′:5′,2″-terselenophene into itsamino analog can moderately improve its water solubility. However, theamino analog is less stable. The amino analog may be transformed intoits γ-glutamate prodrug (as shown in Scheme 9) to further enhance itswater solubility and stability. This conjugate may also enhance targetselectivity for the treatment of kidney cancer because of the higherγ-glutamyl transpeptidase activity in kidney. A modified procedure canalso be designed for the preparation of glutathione conjugate.

[0208] c. Formation of Inclusion Complexes

[0209] The hydrophobic cavity of cyclodextrin derivatives can formstable inclusion complexes with 2-aminomethyl substituted thiophenecompounds. β-Cyclodextrin (cyclic heptaamylose) derivatives are commonlyused for improving water solubility because of their low costs. It isanticipated that the selenophene compounds of the present invention canbe complexed with β-hydroxypropyl, dimethyl and sulfated β-cyclodextrinsto enhance the water solubility of those compounds.

EXAMPLE 53

[0210] Additional National Cancer Institute data demonstratingselenophene growth inhibition of human cancer cell lines is representedin the following tables. The compound must exhibit a Log₁₀ GI50 value of<−4.00 to be considered active against the tested cell line.

NSC: 688829

[0211]

Panel/Cell Line Log₁₀ GI50 Log₁₀ TGI Log₁₀ LC50 Leukemia CCRF-CEM−5.10 >−4.00 >−4.00 K-562 −6.60 >−4.00 >−4.00 MOLT-4 −4.24 >−4.00 >−4.00RPMI-8226 −4.41 >−4.00 >−4.00 SR −4.61 −4.14 >−4.00 Non-Small Cell LungCancer A549/ATCC >−4.00 >−4.00 EKVX >−4.00 >−4.00 >−4.00 HOP-62 −6.60−4.80 >−4.00 HOP-92 −4.01 >−4.00 >−4.00 NCI-H226 −6.05 >−4.00 >−4.00NCI-H23 >−4.00 >−4.00 >−4.00 NCI-H322M −4.56 −4.05 >−4.00 NCI-H460−6.85 >−4.00 >−4.00 NCI-H522 −4.95 −4.40 >−4.00 Colon Cancer COLO 205−4.92 −4.48 −4.04 HCC-2998 −6.41 >−4.00 >−4.00 HCT-116−6.53 >−4.00 >−4.00 HCT-15 −4.64 >−4.00 >−4.00 HT29 −4.63 >−4.00 >−4.00KM12 >−4.00 >−4.00 SW-620 −6.08 >−4.00 >−4.00 CNS CancerSF-295 >−4.00 >−4.00 >−4.00 SF-539 −4.56 >−4.00 >−4.00SNB-19 >−4.00 >−4.00 >−4.00 SNB-75 −4.12 >−4.00 >−4.00 U251 −6.61−4.59 >−4.00 Melanoma LOXIMVI −4.78 >−4.00 >−4.00 MALME-3M−4.54 >−4.00 >−4.00 M14 −4.70 >−4.00 >−4.00 SK-MEL-2 −4.47 >−4.00 >−4.00SK-MEL-28 −4.18 >−4.00 >−4.00 SK-MEL-5 −4.63 >−4.00 >−4.00 UACC-257−6.71 −6.29 >−4.00 UACC-62 −6.85 >−4.00 >−4.00 Ovarian Cancer IGROV1−6.75 −5.61 −4.04 OVCAR-3 −6.89 −6.20 >−4.00 OVCAR-4 −6.73 >−4.00OVCAR-5 −6.91 −6.30 >−4.00 OVCAR-8 −4.82 −4.03 >−4.00 SK-OV-3−4.58 >−4.00 >−4.00 Renal Cancer 786-0 −4.57 −4.19 A498 −7.67 −7.10−6.48 ACHN >−4.00 >−4.00 >−4.00 CAKI-1 −6.72 −6.30 −4.53 SN12C −4.55−4.07 >−4.00 TK-10 −7.55 −6.68 −4.21 UO-31 >−4.00 >−4.00 >−4.00 ProstateCancer PC-3 −4.55 >−4.00 >−4.00 DU-145 >−4.00 >−4.00 >−4.00 BreastCancer MCF7 −6.72 >−4.00 >−4.00 MCF7/ADR-RES −4.52 >−4.00 >−4.00MDA-MB-231/ATCC −4.63 −4.17 >−4.00 HS 578T −5.54 >−4.00 >−4.00MDA-MB-435 −4.48 >−4.00 >−4.00 MDA-N −4.68 >−4.00 >−4.00 BT-549−4.25 >−4.00 >−4.00 T-47D −6.47 >−4.00 >−4.00 MG_MID −5.27 −4.35 −4.06Delta 2.40 2.75 2.42 Range 3.67 3.10 2.48

NSC: 688830

[0212]

Panel/Cell Line Log₁₀ GI50 Log₁₀ TGI Log₁₀ LC50 Leukemia CCRF-CEM−4.59 >−4.00 >−4.00 K-562 −7.14 −4.76 −4.01 MOLT-4 −4.41 >−4.00 >−4.00RPMI-8226 −4.43 >−4.00 >−4.00 SR −4.61 −4.14 >−4.00 Non-Small Cell LungCancer • A549/ATCC −5.22 −4.48 >−4.00 EKVX −4.71 −4.18 >−4.00 HOP-62−7.21 −4.55 −4.07 HOP-92 −4.55 >−4.00 >−4.00 NCI-H322M −7.22 >−4.00NCI-H23 −4.07 >−4.00 >−4.00 NCI-H322M −5.47 −4.70 >−4.00 NCI-H460 −7.77−6.19 −4.54 NCI-H522 >−4.00 >−4.00 >−4.00 Colon Cancer COLO 205 −6.35−5.26 −4.21 HCC-2998 −7.20 −4.94 −4.35 HCT-116 −7.56 −4.76 −4.09 HCT-15−4.54 >−4.00 >−4.00 HT29 −6.65 −4.36 >−4.00 KM12 −4.32 >−4.00 >−4.00SW-620 −7.40 −4.60 >−4.00 CNS Cancer SF-295 −4.68 −4.25 >−4.00 SF-539−4.81 −4.44 −4.08 SNB-19 −4.47 −4.06 >−4.00 SNB-75 −4.59 >−4.00 >−4.00U251 −7.08 −4.68 −4.14 Melanoma LOXIMVI −4.87 −4.56 −4.25 MALME-3M−4.36 >−4.00 >−4.00 M14 −4.43 >−4.00 >−4.00 SK-MEL-2 −4.40 >−4.00 >−4.00SK-MEL-28 >−4.00 >−4.00 >−4.00 SK-MEL-5 −4.44 >−4.00 >−4.00 UACC-257−7.78 −7.34 −6.62 UACC-62 −7.87 >−4.00 >−4.00 Ovarian Cancer IGROV1-−7.85 −6.76 −4.55 OVCAR-3 <−8.00 −7.25 −4.30 OVCAR-4 −6.98 −4.09 OVCAR-5−7.64 >−4.00 OVCAR-8 −5.16 −4.51 >−4.00 SK-OV-3 −5.49 −4.71 −4.29 RenalCancer 786-0 −5.19 −4.64 −4.24 A498 <−8.00 −7.61 −7.14 ACHN−4.68 >−4.00 >−4.00 CAKI-1 <−8.00 −7.55 SN12C −4.31 >−4.00 >−4.00 TK-10<−8.00 −7.43 −4.19 UO-31 >−4.00 >−4.00 >−4.00 Prostate Cancer PC-3−4.51 >−4.00 >−4.00 DU-145 −4.64 >−4.00 >−4.00 Breast Cancer MCF7−7.78 >−4.00 >−4.00 MCF7/ADR-RES −4.91 >−4.00 >−4.00 MDA-MB-231/ATCC−4.90 −4.45 −4.01 HS 578T >−4.00 >−4.00 MDA-MB-435 >−4.00 >−4.00 >−4.00MDA-N −4.01 >−4.00 >−4.00 BT-549 >−4.00 >−4.00 >−4.00 T-47D −6.54 −4.24MG_MID −5.64 −4.62 −4.16 Delta 2.36 2.99 2.98 Range 4.00 3.61 3.14

NSC: 676631

[0213]

Panel/Cell Line Log₁₀GI5O Log₁₀TGI Log₁₀ LC50 LeukemiaCCRF-CEM >−4.00 >−4.00 >−4.00 HL-60(TB) >−4.00 >−4.00 >−4.00 K-562−6.99 >−4.00 >−4.00 MOLT-4 >−4.00 >−4.00 >−4.00RPMI-8226 >−4.00 >−4.00 >−4.00 SR >−4.00 >−4.00 >−4.00 Non-Small CellLung Cancer A549/ATCC −4.23 >−4.00 >−4.00 EKVX >−4.00 >−4.00 >−4.00HOP-62 >−4.00 >−4.00 HOP-92 >−4.00 >−4.00 >−4.00 NCI-H226 −7.30 −6.74−6.28 NCI-H23 −4.78 −4.41 −4.04 NCI-H322M −4.66 >−4.00 >−4.00NCI-H460 >−4.00 >−4.00 NCI-H522 −5.28 −4.67 −4.26 Colon Cancer COLO 205−6.54 −4.90 −4.18 HCC-2998 −4.37 >−4.00 >−4.00 HCT-116−7.27 >−4.00 >−4.00 HCT-15 −4.56 >−4.00 >−4.00 HT29 >−4.00 >−4.00 KM12−4.57 >−4.00 >−4.00 SW-620 −6.45 >−4.00 >−4.00 CNS Cancer SF-268 −4.63−4.19 >−4.00 SF-295 −4.26 >−4.00 >−4.00 SF-539 −4.91 −4.24 >−4.00 SNB-19−4.76 >−4.00 >−4.00 SNB-75 −4.38 >−4.00 >−4.00 U251 −7.11 −4.70 −4.35Melanoma LOX IMVI −4.82 −4.44 −4.07 MALME-3M >−4.00 >−4.00 >−4.00 M14−4.70 >−4.00 >−4.00 SK-MEL-2 >−4.00 >−4.00 >−4.00 SK-MEL-28−4.18 >−4.00 >−4.00 SK-MEL-5 >−4.00 >−4.00 >−4.00 UACC-257 −7.50 >−4.00−6.34 UACC-62 −7.58 −6.94 >−4.00 Ovarian Cancer IGROV1 −7.09 −6.22 −4.29OVCAR-3 −7.56 −6.86 −4.39 OVCAR-4 >−4.00 >−4.00 OVCAR-5 −7.50 −6.68OVCAR-8 −4.61 >−4.00 >−4.00 SK-OV-3 −4.34 >−4.00 >−4.00 Renal Cancer786-0 >−4.00 >−4.00 >−4.00 A498 −7.56 −7.08 −6.52ACHN >−4.00 >−4.00 >−4.00 CAKI-1 −7.51 −6.72 −4.19 RXF-393−4.14 >−4.00 >−4.00 SN12C >−4.00 >−4.00 >−4.00 TK-10 −7.30 −6.43 −4.14UO-31 >−4.00 >−4.00 >−4.00 Prostate Cancer PC-3 −4.28 >−4.00 >−4.00DU-145 −4.70 −4.11 >−4.00 Breast Cancer MCF7 −7.03 −4.66 >−4.00MCF7/ADR-RES −5.00 −4.50 −4.01 MDA-MB-231/ATCC −4.72 −4.06 >−4.00 HS578T >−4.00 >−4.00 >−4.00 MDA-MB-435 −5.09 −4.26 >−4.00 MDA-N −4.78−4.36 >−4.00 BT-549 −4.77 −4.42 −4.06 T-47D −6.17 >−4.00 >−4.00 MG_MID−5.18 −4.47 −4.15 Delta 2.40 2.61 2.37 Range 3.58 3.08 2.52

NSC: 675246

[0214]

Panel/Cell Line Log₁₀GI50 Log₁₀ TGI Log₁₀ LC50 Leukemia CCRF-CEM −5.50−4.91 >−4.00 HL-60(TB) −5.24 >−4.00 >−4.00 K-562 −6.43 −5.12 >−4.00MOLT-4 −5.49 −4.92 >−4.00 RPMI-8226 −5.13 >−4.00 >−4.00 SR−5.18 >−4.00 >−4.00 Non-Small Cell Lung Cancer A549/ATCC −4.98 −4.65−4.33 EKVX −5.26 −4.76 −4.37 HOP-62 −5.14 −4.69 −4.35 HOP-92 −5.58 −4.95−4.42 NCI-H322M −6.21 −5.63 NCI-H23 −4.88 −4.51 −4.15 NCI-H322M −4.97−4.65 −4.32 NCI-H460 −6.49 −5.44 −4.64 NCI-H52 −5.61 −5.18 −4.63 ColonCancer COLO 205 −6.00 −5.39 >−4.00 HCC-2998 −5.91 −4.93 −4.27 HCT-116−7.19 −4.97 −4.49 HCT-15 −5.37 −4.75 >−4.00 HT29 −6.07 −4.97 −4.23 KM12−5.38 −4.77 −4.28 SW-620 −6.38 −4.99 −4.45 CNS Cancer SF-268−5.39 >−4.00 >−4.00 SF-295 −4.97 −4.56 −4.14 SF-539 −4.91 −4.60 −4.29SNB-19 −4.91 −4.28 >−4.00 SNB-75 −5.41 >−4.00 >−4.00 U251 −7.15 −4.91−4.26 Melanoma LOXIMVI −5.44 −5.01 −4.43 MALME-3M −5.21 −4.71 −4.30 M14−5.06 −4.62 −4.21 SK-MEL-2 −5.04 −4.59 −4.16 SK-MEL-28 −5.13 −4.66 −4.27SK-MEL-5 −5.58 −5.05 −4.53 UACC-257 −7.30 −6.65 UACC-62 −7.99 −4.95−4.17 Ovarian Cancer IGROV1 −7.27 −5.65 −4.91 OVCAR-3 −7.22 −5.89 −5.16OVCAR-4 −6.24 −4.91 −4.32 OVCAR-5 −6.74 −4.60 >−4.00 OVCAR-8 −5.30−4.48 >−4.00 Renal Cancer 786-0 −5.66 −5.28 −4.76 A498 −7.41 −6.78 −6.16CAKI-1 −5.27 −4.73 −4.29 RXF-393 −7.68 −7.04 TK-10 −5.72 −4.97 >−4.00UO-31 −5.21 −4.41 >−4.00 −7.50 −6.65 −4.05 −4.92 −4.61 −4.29 ProstateCancer PC-3 −5.42 −4.88 −4.44 DU-145 −4.99 −4.66 −4.33 Breast CancerMCF7 −6.84 −5.42 −4.53 MCF7/ADR-RES −5.29 −4.33 >−4.00 MDA-MB-231/ATCC−5.36 −4.47 >−4.00 MDA-N −5.01 −4.25 >−4.00 T-47D −5.42 −4.82 −4.35−5.56 −4.90 −4.40 −5.20 −4.68 −4.26 −5.63 −4.33 >−4.00 MG_MID −5.78−4.91 −4.29 Delta 2.22 2.14 1.88 Range 3.11 3.04 2.16

NSC: 675247

[0215]

Panel/Cell Line Log₁₀ GI50 Log₁₀ TGI Log₁₀ LC50 Leukemia CCRF-CEM −5.47−5.00 >−4.00 HL-60(TB) −5.39 K-562 −5.88 −5.30 −4.15 MOLT-4 −5.48−5.08 >−4.00 RPMI-8226 −5.39 >−4.00 >−4.00 SR −5.43 >−4.00 >−4.00 >−4.00Non-Small Cell Lung Cancer A549/ATCC −5.34 −4.71 −4.27 EKVX −5.30−4.46 >−4.00 HOP-62 −5.26 −4.67 −4.24 HOP-92 −5.62 −5.12 −4.26 NCI-H226−5.77 −5.41 −5.04 NCI-H23 −5.18 −4.60 −4.02 NCI-H322M −4.95 −4.61 −4.28NCI-H460 −6.28 −5.00 −4.09 NCI-H522 −5.78 −5.47 −5.15 Colon Cancer COLO205 −5.71 −5.29 −4.75 HCC-2998 −6.07 −5.49 −4.90 HCT-116 −6.27−4.95 >−4.00 HCT-15 −5.42 −4.91 −4.26 HT29 −5.79 −5.12 −4.18 KM12 −5.35−4.82 −4.30 SW-620 −5.87 −5.44 −5.02 CNS Cancer SF-268 −5.76 −5.36 −4.68SF-295 −4.93 −4.54 −4.15 SF-539 −4.76 −4.39 −4.02 SNB-19 −5.26 −4.60−4.04 SNB-75 −4.83 −4.41 >−4.00 U251 −6.33 −4.91 −4.30 Melanoma LOXIMVI−5.52 −5.15 >−4.00 MALME-3M −5.54 −4.97 −4.25 M14 −5.43 −4.89 −4.31SK-MEL-2 −5.15 −4.62 −4.16 SK-MEL-28 −5.29 >−4.00 >−4.00 SK-MEL-5 −5.84−5.52 −5.20 UACC-257 −6.36 −5.70 −4.50 UACC-62 −6.81 −5.61 −4.35 OvarianCancer IGROV1 −5.86 −5.19 −4.52 OVCAR-3 −6.68 −5.93 −5.27 OVCAR-4 −5.77−5.16 −4.31 OVCAR-5 −5.88 −4.89 >−4.00 OVCAR-8 −5.43 −4.75 −4.14 RenalCancer 786-0 −5.56 −5.20 −4.15 A498 −6.42 −5.85 −5.14 ACHN−5.40 >−4.00 >−4.00 CAKI-1 −7.13 −6.33 −4.88 RXF-393 −5.80 −5.43 −5.06SN12C −5.56 −4.93 >−4.00 TK-10 −6.86 −6.18 −4.80 UO-31 −5.28 −4.83 −4.41Prostate Cancer PC-3 −5.45 −4.91 −4.45 DU-145 −5.16 −4.71 −4.33 BreastCancer MCF7 −6.80 −4.95 −4.32 MCF7/ADR-RES −5.42 −4.58 >−4.00 MDA-MB-−5.30 −4.79 −4.32 3221/ATCC −5.27 −4.44 >−4.00 HS 578T −5.49 −4.95 −4.33MDA-MB-435 −5.53 −5.01 −4.31 MDA-N −5.11 −4.60 −4.13 BT-549 −5.47−4.85 >−4.00 T-47D MG_MID −5.65 −4.98 −4.34 Delta 1.48 1.36 0.93 Range2.37 2.33 1.27

NSC: 675343

[0216]

Panel/Cell Line Log₁₀ GI50 Log₁₀ TGI Log₁₀ LC50 Leukemia CCRF-CEM −4.65−4.22 >−4.00 HL-60(TB) −4.21 >−4.00 >−4.00 K-562 <−8.00 −4.58 >−4.00MOLT-4 −4.54 −4.06 >−4.00 RPMI-8226 >−4.00 >−4.00 >−4.00SR >−4.00 >−4.00 >−4.00 Non-Small Cell Lung CancerA549/ATCC >−4.00 >−4.00 >−4.00 EKVX >−4.00 >−4.00 >−4.00HOP-62 >−4.00 >−4.00 >−4.00 HOP-92 −4.39 >−4.00 >−4.00 NCI-H226 <−8.00−7.14 >−4.00 NCI-H23 >−4.00 >−4.00 >−4.00 NCI-H322M −5.01 >−4.00 >−4.00NCI-H460 <−8.00 >−4.00 >−4.00 NCI-H522 −4.00 >−4.00 >−4.00 Colon CancerCOLO 205 <−8.00 −5.98 −4.64 HCC-2998 <−8.00 −5.47 >−4.00 HCT-116<−8.00 >−4.00 >−4.00 HCT-15 >−4.00 >−4.00 >−4.00 HT29 >−4.00 >−4.00KM12 >−4.00 >−4.00 >−4.00 SW-620 <−8.00 >−4.00 >−4.00 CNS CancerSF-268 >−4.00 >−4.00 >−4.00 SF-295 >−4.00 >−4.00 >−4.00SF-539 >−4.00 >−4.00 >−4.00 SNB-19 >−4.00 >−4.00 >−4.00 SNB-75 U251<−8.00 >−4.00 >−4.00 Melanoma LOX IMVI >−4.00 >−4.00 >−4.00MALME-3M >−4.00 >−4.00 >−4.00 M14 >−4.00 >−4.00 >−4.00SK-MEL-2 >−4.00 >−4.00 >−4.00 SK-MEL-28 >−4.00 >−4.00 >−4.00SK-MEL-5 >−4.00 >−4.00 UACC-257 <−8.00 <−8.00 UACC-62<−8.00 >−4.00 >−4.00 Ovarian Cancer OVCAR-3 −7.59 −4.49 >−4.00 OVCAR-4<−8.00 >−4.00 >−4.00 OVCAR-5 −6.80 >−4.00 >−4.00 OVCAR-8<−8.00 >−4.00 >−4.00 SK-OV-3 >−4.00 >−4.00 >−4.00 Renal Cancer786-0 >−4.00 >−4.00 >−4.00 A498 <−8.00 <−8.00 −6.80ACHN >−4.00 >−4.00 >−4.00 CAKI-1 <−8.00 <−8.00 >−4.00 RXF-393−4.68 >−4.00 >−4.00 SN12C >−4.00 >−4.00 >−4.00 TK-10 <−8.00<−8.00 >−4.00 UO-31 >−4.00 >−4.00 >−4.00 Prostate CancerPC-3 >−4.00 >−4.00 >−4.00 DU-145 >−4.00 >−4.00 >−4.00 Breast Cancer MCF7<−8.00 >−4.00 >−4.00 MCF7/ADR-RES >−4.00 >−4.00 >−4.00MDA-MB-231/ATCC >−4.00 >−4.00 >−4.00 HS 578T >−4.00 >−4.00 >−4.00MDA-MB-435 >−4.00 >−4.00 >−4.00 MDA-N >−4.00 >−4.00 >−4.00BT-549 >−4.00 >−4.00 >−4.00 T-47D −6.14 >−4.00 >−4.00 MG_MID Delta −5.36−4.41 −4.06 Range 2.64 3.59 2.74 4.00 4.00 2.80

NSC: 676632

[0217]

Panel/Cell Line Log₁₀ GI50 Log₁₀ TGI Log₁₀LC50 Leukemia CCRF-CEM−4.06 >−4.00 >−4.00 HL-60(TB) >−4.00 >−4.00 >−4.00 K-562−7.32 >−4.00 >−4.00 MOLT-4 >−4.00 >−4.00 >−4.00RPM1-8226 >−4.00 >−4.00 >−4.00 SR >−4.00 >−4.00 >−4.00 Non-Small CellLung Cancer A549/ATCC −5.58 >−4.00 >−4.00 EKVX −4.30 >−4.00 >−4.00HOP-62 −7.12 >−4.00 >−4.00 HOP-92 >−4.00 >−4.00 >−4.00 NCI-H226 −7.71−7.27 −6.49 NCI-H23 >−4.00 >−4.00 >−4.00 NCI-H322M >−4.00 >−4.00 >−4.00NCI-H460 −7.33 >−4.00 >−4.00 NCI-H522 >−4.00 >−4.00 >−4.00 Colon CancerHCC-2998 −6.75 −6.26 −5.61 HCT-116 >−4.00 >−4.00 >−4.00 HCT-15−7.35 >−4.00 >−4.00 HT29 >−4.00 >−4.00 >−4.00 KM12 −6.27 >−4.00 >−4.00SW-620 >−4.00 >−4.00 >−4.00 −6.82 >−4.00 >−4.00 CNS CancerSF-268 >−4.00 >−4.00 >−4.00 SF-295 >−4.00 >−4.00 >−4.00SF-539 >−4.00 >−4.00 >−4.00 SNB-19 −4.37 >−4.00 >−4.00SNB-75 >−4.00 >−4.00 >−4.00 U251 −7.45 −4.92 −4.33 Melanoma LOXIMVI >−4.00 <−4.00 >−4.00 MALME-3M >−4.00 >−4.00 >−4.00M14 >−4.00 >−4.00 >−4.00 SK-MEL-2 >−4.00 >−4.00 >−4.00SK-MEL-28 >−4.00 >−4.00 >−4.00 SK-MEL-5 −4.20 >−4.00 >−4.00 UACC-257−7.67 >−4.00 UACC-62 −7.65 −7.25 >−4.00 Ovarian Cancer IGROV1 −7.49−6.56 >−4.00 OVCAR-3 −7.71 −7.17 >−4.00 OVCAR-4 −6.87 >−4.00 OVCAR-5−7.88 −7.11 −6.11 OVCAR-8 >−4.00 >−4.00 >−4.00 >−4.00 >−4.00 >−4.00Renal Cancer 786-0 >−4.00 >−4.00 >−4.00 A498 −7.73 −7.37 −7.02ACHN >−4.00 >−4.00 >−4.00 CAKI-1 −7.90 −6.91 >−4.00RXF-393 >−4.00 >−4.00 >−4.00 SN12C >−4.00 >−4.00 >−4.00 TK-10 −7.55−7.09 >−4.00 UO-31 >−4.00 >−4.00 >−4.00 Prostate CancerPC-3 >−4.00 >−4.00 >−4.00 DU-145 −4.02 >−4.00 >−4.00 Breast Cancer MCF7−7.91 >−4.00 >−4.00 MCF7/ADR-RES >−4.00 >−4.00 >−4.00MDA-MB- >−4.00 >−4.00 >−4.00 231/ATCC >−4.00 >−4.00 >−4.00HS578T >−4.00 >−4.00 >−4.00 MDA-MB-435 >−4.00 >−4.00 >−4.00MDA-N >−4.00 >−4.00 >−4.00 BT-549 −6.71 >−4.00 >−4.00 T-47D MG_MID −5.16−4.48 −4.16 Delta 2.75 2.89 2.86 Range 3.91 3.37 3.02

NSC: 675344

[0218]

Panel/Cell Line Log₁₀ GI50 Log₁₀ TGI Log₁₀ LC50 LeukemiaCCRF-CEM >−4.00 >−4.00 >−4.00 HL-60(TB) >−4.00 >−4.00 >−4.00 K-562−7.36 >−4.00 >−4.00 MOLT-4 >−4.00 >−4.00 >−4.00RPMI-8826 >−4.00 >−4.00 >−4.00 SR >−4.00 >−4.00 >−4.00 Non-Small CellLung Cancer A549/ATCC −4.34 >−4.00 >−4.00 EKVX >−4.00 >−4.00 >−4.00HOP-62 −4.46 >−4.00 >−4.00 HOP-92 −4.56 −4.05 >−4.00 NCI-H226 <−8.00<−8.00 −6.65 NCI-H23 −4.69 >−4.00 >−4.00 NCI-H322M −4.68 >−4.00 >−4.00NCI-H460 <−8.00 >−4.00 >−4.00 NCI-H522 >−4.00 >−4.00 >−4.00 Colon CancerHCC-2998 −6.59 −5.79 −5.01 HCT-116 <−8.00 −7.38 −5.53 HCT-15−7.59 >4.00 >−4.00 HT29 >4.00 >−4.00 >−4.00 KM12 >−4.00 >−4.00 >−4.00SW-620 >4.00 >−4.00 >−4.00 −7.06 >−4.00 >−4.00 CNS Cancer SF-268−4.63 >−4.00 >−4.00 SF-295 −4.57 >−4.00 >−4.00SF-539 >−4.00 >−4.00 >−4.00 SNB-19 −4.53 >−4.00 >−4.00 SNB-75 −4.78−4.31 >−4.00 U251 −7.60 −4.58 >−4.00 Melanoma LOX IMVI−4.46 >−4.00 >−4.00 MALME-3M >−4.00 >−4.00 >−4.00M14 >−4.00 >−4.00 >−4.00 SK-MEL-2 >−4.00 >−4.00 >−4.00SK-MEL-28 >−4.00 >−4.00 >−4.00 SK-MEL-5 >−4.00 >−4.00 >−4.00 UACC-257<−8.00 −7.73 −7.28 UACC-62 <−8.00 −7.85 >−4.00 Ovarian Cancer IGROV1−7.91 −7.37 −4.79 OVCAR-3 <−8.00 >−4.00 >−4.00 OVCAR-4−7.38 >−4.00 >−4.00 OVCAR-5 <−8.00 −7.06 >−4.00OVCAR-8 >−4.00 >−4.00 >−4.00 SK-OV-3 −4.78 >−4.00 >−4.00 Renal Cancer786-0 −4.91 −4.17 >−4.00 A498 <−8.00 −7.74 −7.18 ACHN −4.89 −4.08 >−4.00RXF-393 −4.80 −4.28 >−4.00 SN12C >−4.00 >−4.00 >−4.00 TK-10 <−8.00−7.30 >−4.00 UO-31 >−4.00 >−4.00 >−4.00 Prostate CancerPC-3 >−4.00 >−4.00 >−4.00 DU-145 −4.44 >−4.00 >−4.00 Breast Cancer MCF7−6.90 >−4.00 >−4.00 MCF7/ADR-RES >−4.00 >−4.00 >−4.00 MDA-MB-231/ATCC−4.72 >−4.00 >−4.00 MS 578T −4.27 >−4.00 >−4.00MDA-MB-435 >−4.00 >−4.00 >−4.00 MDA-N >−4.00 >−4.00 >−4.00 BT-549−4.37 >−4.00 >−4.00 T-47D −6.21 >−4.00 >−4.00 MG_MID −5.28 −4.54 −4.21Delta 2.72 3.46 3.07 Range 4.00 4.00 3.28

NSC: 676630

[0219]

Panel/Cell Line Log₁₀ GI50 Log₁₀ TGI Log₁₀ LC50 LeukemiaCCRF-CEM >−4.00 >−4.00 >−4.00 HL-60(TB) >−4.00 >−4.00 >−4.00 K-562  −5.47 >−4.00 >−4.00 MOLT-4 >−4.00 >−4.00 >−4.00RPMI-8226 >−4.00 >−4.00 >−4.00 SR >−4.00 >−4.00 >−4.00 Non-Small CellLung Cancer A549/ATCC   −4.06 >−4.00 >−4.00 EKVX >−4.00 >−4.00 >−4.00HOP-62 >−4.00 >−4.00 HOP-92   >4.00 >−4.00 >−4.00 NCI-H226   −6.72  −6.31   −5.61 NCI-H23 >−4.00 >−4.00 NCI-H322M >−4.00 >−4.00 >−4.00NCI-H460   −6.89 >−4.00 >−4.00 NCI-H522   −4.68   −4.27 >−4.00 ColonCancer COLO 205   −4.03 >−4.00 >−4.00 HCC-2998   −4.07 >−4.00 HCT-116  −5.52 >−4.00 >−4.00 HCT-15 >−4.00 >−4.00 >−4.00HT29 >−4.00 >−4.00 >−4.00 KM12 >−4.00 >−4.00 >−4.00SW-620 >−4.00 >−4.00 >−4.00 CNS Cancer SF-268 >−4.00 >−4.00 >−4.00SF-295 >−4.00 >−4.00 >−4.00 SF-539 >−4.00 >−4.00 >−4.00SNB-19 >−4.00 >−4.00 >−4.00 SNB-75   −4.17 >−4.00 >−4.00 U251  −6.23 >−4.00 >−4.00 Melanoma LOC IMVI >−4.00 >−4.00 >−4.00MALME-3M >−4.00 >−4.00 >−4.00 M14 >−4.00 >−4.00 >−4.00SK-MEL-2 >−4.00 >−4.00 >−4.00 SK-MEL-28 >−4.00 >−4.00 >−4.00SK-MEL-5 >−4.00 >−4.00 >−4.00 UACC-257   −6.56   −6.17   −5.16 UACC-62  −6.58 >−4.00 >−4.00 Ovarian Cancer IGROV1   −5.83 >−4.00 >−4.00OVCAR-3   −6.14 >−4.00 >−4.00 OVCAR-4 >−4.00 OVCAR-5   −6.88   −6.35  −5.45 OVCAR-8   −4.48 >−4.00 >−4.00 SK-OV-3 >−4.00 >−4.00 >−4.00 RenalCancer 786-0 >−4.00 >−4.00 >−4.00 ACHN >−4.00 >−4.00 >−4.00 CAKI-1  −6.13   −4.82 >−4.00 RXF-393 >−4.00 >−4.00 >−4.00SN12C >−4.00 >−4.00 >−4.00 TK-10   −6.42   −5.79 >−4.00UO-31 >−4.00 >−4.00 >−4.00 Prostate Cancer PC-3 >−4.00 >−4.00 >−4.00DU-145 >−4.00 >−4.00 >−4.00 Breast Cancer MCF7   −6.17 >−4.00 >−4.00MCF7/ADR-RES   −4.10 >−4.00 >−4.00 MDA-MB-231/ATCC >−4.00 >−4.00 >−4.00HS 578T >−4.00 >−4.00 >−4.00 MDA-MB-435 >−4.00 >−4.00 >−4.00MDA-N >−4.00 >−4.00 >−4.00 BT-549   −4.20 >−4.00 >−4.00T-47D >−4.00 >−4.00 MG_MID Delta   −4.58   −4.17   −4.07 Range   2.31   2.18    1.54    2.89    2.35    1.61 

NSC: 675245

[0220]

Panel/Cell Line Log₁₀ GI50 Log₁₀ TGI Log₁₀ LC50 LeukemiaCCRF-CEM >−4.00 >−4.00 >−4.00 HL-60(TB) >−4.00 >−4.00 >−4.00 K-562  −5.17 >−4.00 >−4.00 MOLT-4 >−4.00 >−4.00 >−4.00RPMI-8226 >−4.00 >−4.00 >−4.00 SR >−4.00 >−4.00  >−4.00 Non-Small CellLung Cancer A549/ATCC >−4.00 >−4.00 >−4.00 EKVX >−4.00 >−4.00 >−4.00HOP-62   −4.29 >−4.00 >−4.00 HOP-92 >−4.00 >−4.00 >−4.00 NCI-H226  −6.01   −5.44 >−4.00 NCI-H23 >−4.00 >−4.00 >−4.00NCI-H322M >−4.00 >−4.00 >−4.00 NCI-H460   −5.75 >−4.00 >−4.00 NCI-H522  −4.66   −4.34   −4.02 Colon Cancer COLO 205 >−4.00 >−4.00 >−4.00HCC-2998   −5.32 >−4.00 >−4.00 HCT-116   −6.16 >−4.00 >−4.00HCT-15 >−4.00 >−4.00 >−4.00 HT29 >−4.00 >−4.00 >−4.00KM12 >−4.00 >−4.00 >−4.00 SW-620   −5.38 >−4.00 >−4.00 CNS CancerSF-268 >−4.00 >−4.00 >−4.00 SF-295   −4.31 >−4.00 >−4.00SF-539 >−4.00 >−4.00 >−4.00 SNB-19   −4.53 >−4.00 >−4.00 SNB-75  −5.00 >−4.00 >−4.00 U251   −5.98 >−4.00 >−4.00 Melanoma LOXIMVI >−4.00 >−4.00 >−4.00 MALME-3M >−4.00 >−4.00 >−4.00M14 >−4.00 >−4.00 >−4.00 SK-MEL-2 >−4.00 >−4.00 >−4.00SK-MEL-28 >−4.00 >−4.00 >−4.00 SK-MEL-5   −4.14 >−4.00 >−4.00 UACC-257  −6.49   −6.04 >−4.00 UACC-62   −7.19 >−4.00 >−4.00 Ovarian CancerIGROV1   −4.17   −4.59   −4.13 OVCAR-3   −6.41 >−4.00 >−4.00 OVCAR-4  −5.58 >−4.00 >−4.00 OVCAR-5   −6.11 >−4.00 >−4.00OVCAR-8 >−4.00 >−4.00 >−4.00 Renal Cancer 786-0   −4.17 >−4.00 >−4.00A498   −6.51   −6.05   −5.17 ACHN   −4.24 >−4.00 >−4.00 CAKI-1   −6.57  −6.03 >−4.00 RXF-393   −4.92 >−4.00 >−4.00 SN12C >−4.00 >−4.00 >−4.00TK-10   −6.59   −6.09 >−4.00 UO-31 >−4.00 >−4.00 >−4.00 Prostate CancerPC-3 >−4.00 >−4.00 >−4.00 DU-145 >−4.00 >−4.00 >−4.00 Breast Cancer MCF7  −5.83 >−4.00 >−4.00 MCF7/ADR-RES >−4.00 >−4.00 >−4.00MDA-MB-231/ATCC >−4.00 >−4.00 >−4.00 HS 578T   −4.06 >−4.00 >−4.00MDA-MB-435 >−4.00 >−4.00 >−4.00 MDA-N >−4.00 >−4.00 >−4.00BT-549 >−4.00 >−4.00 >−4.00 T-47D   −4.80 >−4.00 >−4.00 MG_MID Delta  −4.67   −4.18   −4.02 Range   2.53    1.91    1.15    3.19    2.09   1.17 

NSC: 675244

[0221]

Panel/Cell Line Log₁₀ GI50 Log₁₀ TGI Log₁₀ LC50 Leukemia CCRF-CEM −5.67−5.15 >−4.00   HL-60(TB) −5.61 >−4.00   K-562 −5.87 −4.48 >−4.00  MOLT-4 −5.62 −5.12 >−4.00   RPMI-8226 −5.53 >−4.00   >−4.00   −5.39Non-Small Cell Lung Cancer A549/ATCC −4.87 −4.58 −4.29 EKVX −4.60−4.19 >−4.00   HOP-62 −4.92 −4.53 −4.13 HOP-92 −4.96 −4.59 −4.20NCI-H226 −5.65 −5.29 −4.41 NCI-H23 −4.87 −4.51 −4.16 NCI-H322M −4.88−4.55 −4.22 NCI-H460 −5.65 −4.79 −4.39 NCI-H522 −5.28 −4.76 −4.37 ColonCancer COLO 205 −5.50 −4.92 −4.46 HCC-2998 −5.60 −4.94 −4.41 HCT-116−6.42 −4.86 −4.41 HCT-15 −5.13 −4.58 −4.10 HT29 −5.49 −4.88 −4.44 KM12−5.20 −4.71 −4.34 SW-620 CNS Cancer SF-268 −5.35 −4.24 >−4.00   SF-295−4.82 −4.55 −4.27 SF-539 −4.77 −4.48 −4.20 SNB-19 −4.99 −4.52 −4.06SNB-75 −5.70 −5.19 >−4.00   U251 −6.50 −4.86 −4.32 Melanoma LOX IMVI−5.25 −4.68 −4.11 MALME-3M −4.90 −4.55 −4.19 M14 −4.94 −4.54 −4.14SK-MEL-2 −4.78 −4.34 >−4.00   SK-MEL-28 −4.88 −4.57 −4.26 SK-MEL-5 −5.47−4.86 −4.43 UACC-257 −6.51 −6.07 −4.48 UACC-62 −7.15 −4.97 −4.18 OvarianCancer IGROV1 −5.48 −4.81 −4.40 OVCAR-3 −6.55 −5.10 −4.51 OVCAR-4 −5.80−4.83 −4.22 OVCAR-5 −6.26 −4.88 −4.12 OVCAR-8 −5.04 −4.52 −4.02 RenalCancer 786-0 −5.22 −4.73 −4.36 A8498 −5.85 −5.53 −5.20 ACHN −4.99 −4.66−4.33 CAKI-1 −6.69 −6.15 −4.84 RXF-393 −5.71 −5.09 >−4.00   SN12C −5.00−4.65 −4.30 TK-10 −6.54 −6.05 −4.60 UO-31 −4.81 −4.54 −4.27 ProstateCancer PC-3 −4.92 −4.49 −4.06 DU-145 −4.90 −4.60 −4.29 Breast CancerMCF7 −6.32 −4.94 −4.36 MCF7/ADR-RES HS 578T −5.14 −4.66 −4.27 MDA-MB-435−4.99 −4.48 >−4.00   MDA-N −5.34 −4.71 −4.19 BT-549 −4.91 −4.51 −4.10T-47D −4.95 −4.63 −4.32 −5.42 −4.45 >−4.00   MG_MID Delta −5.43 −4.78−4.25 Range   1.72   1.37   0.96   2.55   2.15   1.20

NSC: 675346

[0222]

Panel/Cell Line Log₁₀ GI50 Log₁₀ TGI Log₁₀ LC50 LeukemiaCCRF-CEM >−4.00   >−4.00 >−4.00 HL-60(TB) >−4.00   >−4.00 >−4.00 K-562−5.64 >−4.00 >−4.00 MOLT-4 >−4.00   >−4.00 >−4.00RPMI-8226 >−4.00   >−4.00 >−4.00 SR >−4.00   >−4.00 >−4.00 Non-SmallCell Lung Cancer A549/ATCC −4.26 >−4.00 >−4.00 EKVX −4.11 >−4.00 >−4.00HOP-62 −4.19 >−4.00 >−4.00 HOP-92 −4.73   −4.15 >−4.00 NCI-H226 −7.76  −6.92   −5.98 NCI-H23 −4.84   −4.23 >−4.00 NCI-H322M−4.89 >−4.00 >−4.00 NCI-H460 −6.60 >−4.00 >−4.00 NCI-H522−4.48 >−4.00 >−4.00 Colon Cancer COLO 205 −5.32 >−4.00 >−4.00 HCC-2998−6.63   6.15    −4.68 HCT-116 −6.94 >−4.00 >−4.00 HCT-15−4.73 >−4.00 >−4.00 HT29 >−4.00   >−4.00 >−4.00 KM12 −4.49 >−4.00 >−4.00SW-620 >−4.00 >−4.00 >−4.00 >−4.00 CNS Cancer SF-268 −4.72 >−4.00 >−4.00SF-295 −4.67   −4.16 >−4.00 SF-539 −4.29 >−4.00 >−4.00 SNB-19 −4.66  −4.02 >−4.00 SNB-75 −4.86   −4.24 >−4.00 U251 −7.24   −4.59 >−4.00Melanoma LOX IMVI −4.71 >−4.00 >−4.00 MALME-3M −4.46 >−4.00 >−4.00 M14−4.55 >−4.00 >−4.00 SK-MEL-2 −4.61 >−4.00 >−4.00 SK-MEL-28−4.35 >−4.00 >−4.00 SK-MEL-5 −4.33 >−4.00 >−4.00 UACC-257 −7.58   −7.08  −6.51 UACC-62 <−8.00     −7.55   −4.07 Ovarian Cancer IGROV1 −6.79  −6.25 >−4.00 OVCAR-3 −7.72   −4.67 >−4.00 OVCAR-4 −6.92   −4.57 >−4.00OVCAR-5 −7.35   −6.24 >−4.00 OVCAR-8 −5.35 >−4.00 >−4.00 SK-OV-3−4.90 >−4.00 >−4.00 Renal Cancer 786-0 −4.87 >−4.00 >−4.00 A498 −6.84  −6.53   −6.03 ACHN −4.64 >−4.00 >−4.00 RXF-393 −4.79   −4.37 >−4.00SN12C >−4.00   >−4.00 >−4.00 TK-10 −7.20   −6.38   −4.27 UO-31−4.45 >−4.00 >−4.00 Prostate Cancer PC-3 −4.21 >−4.00 >−4.00 DU-145−4.54 >−4.00 >−4.00 Breast Cancer MCF7 −6.09 >−4.00 >−4.00 MCF7/ADR-RES−4.88   −4.22 >−4.00 HS 578T −4.63 >−4.00 >−4.00 MDA-MB-435 −4.55  −4.06 >−4.00 MDA-N >−4.00   >−4.00 >−4.00 BT-549 −4.00 >−4.00 >−4.00T-47D −4.74   −4.39   −4.04 −5.77 >−4.00 >−4.00 MG_MID Delta −5.17  −4.42   −4.13 Range   2.83   3.13    2.38    4.00   3.55    2.51 

NSC: 675345

[0223]

Panel/Cell Line Log₁₀ GI50 Log₁₀ TGI Log₁₀ LC50 LeukemiaCCRF-CEM >−4.00   >−4.00 >−4.00 HL-60(TB) >−4.00   >−4.00 >−4.00 K-562−4.51 >−4.00 >−4.00 MOLT-4 >−4.00   >−4.00 >−4.00RPMI-8226 >−4.00   >−4.00 >−4.00 SR >−4.00   >−4.00 >−4.00 Non-SmallCell Lung Cancer A549/ATCC >−4.00   >−4.00 >−4.00EKVX >−4.00   >−4.00 >−4.00 HOP-62 −4.32 >−4.00 >−4.00 HOP-92 −4.84  −4.26 >−4.00 NCI-H226 −6.24   −5.61   −5.07 NCI-H23−4.66 >−4.00 >−4.00 NCI-H322M >−4.00   >−4.00 >−4.00 NCI-H460−6.09 >−4.00 >−4.00 NCI-H522 −4.07 >−4.00 >−4.00 Colon Cancer COLO 205−4.47 >−4.00 >−4.00 HCC-2998 −5.63   −5.16   −4.13 HCT-116−5.39 >−4.00 >−4.00 HCT-15 >−4.00   >−4.00 >−4.00HT29 >−4.00   >−4.00 >−4.00 KM12 >−4.00   >−4.00 >−4.00 SW-620−4.42 >−4.00 >−4.00 CNS Cancer SF-268 >−4.00   >−4.00 >−4.00 SF-295−4.41 >−4.00 >−4.00 SF-539 >−4.00   >−4.00 >−4.00 SNB-19−4.50 >−4.00 >−4.00 SNB-75 −4.28 >−4.00 >−4.00 U251 −5.64   −4.67  −4.05 Melanoma LOX IMVI >−4.00   >−4.00 >−4.00MALME-3M >−4.00   >−4.00 >−4.00 M14 >−4.00   >−4.00 >−4.00 SK-MEL-2−4.11 >−4.00 >−4.00 SK-MEL-28 >−4.00   >−4.00 >−4.00SK-MEL-5 >−4.00   >−4.00 >−4.00 UACC-257 −5.86 >−4.00   −5.24 UACC-62−6.46   −5.55 >−4.00 Ovarian Cancer IGROV1 −5.36   −4.64 >−4.00 OVCAR-3−5.66 >−4.00   −5.39 OVCAR-4 >−4.00 >−4.00 OVCAR-5 −5.81   −5.27 >−4.00OVCAR-8 >−4.00   >−4.00 >−4.00 SK-OV-3 −4.26 >−4.00 >−4.00 Renal Cancer786-0 −4.96 >−4.00 >−4.00 A498 −6.32   −5.81   −5.39 ACHN−4.11 >−4.00 >−4.00 CAKI-1 −4.49 >−4.00 >−4.00RXF-393 >−4.00   >−4.00 >−4.00 SN12C −6.59   −5.78 >−4.00TK-10 >−4.00   >−4.00 >−4.00 UO-31 Prostate CancerPC-3 >−4.00   >−4.00 >−4.00 DU-145 >−4.00   >−4.00 >−4.00 Breast CancerMCF7 −6.14   −5.25 >−4.00 MCF7/ADR-RES >−4.00   >−4.00 >−4.00MDA-MB-231/ATC −4.74 >−4.00 >−4.00 HS 578T −4.17 >−4.00 >−4.00MDA-MB-435 −4.23 >−4.00 >−4.00 MDA-N >−4.00   >−4.00 >−4.00BT-549 >−4.00   >−4.00 >−4.00 T-47D −4.10 >−4.00 >−4.00 MG_MID Delta−4.75   −4.21   −4.07 Range   2.03   1.61    1.32    2.59   1.81   1.39 

EXAMPLE 54

[0224] Inhibition of Protein Kinase C

[0225] The Protein Kinase C (PKC) screening assay utilized in thefollowing experiments is similar to standard PKC assays used by manyinvestigators. Its primary features are that 1) the assay utilizes a50:50 mixture of recombinant mouse PKCα and mouse PKCβ₂; 2) employshistone as phosphate-accepting substrate; and 3) the PKC enzymaticactivity is activated with phosphatidylserine, TPA and low concentrationof calcium, so that both calcium and TPA are somewhat limiting for theextent of activation. In this manner the assay is sensitive toinhibitors of PKC activation. A more detailed description of the assayis provided in the following paragraphs.

[0226] The recombinant PKC formulation is a mixture (equal parts byactivity) of mouse PKCα and mouse PKCβ₂. The enzymes are expressed inSf9 insect cells from recombinant baculovirus and partially purified onDEAE-cellulose and Sephacryl 200 gel filtration. Sufficient PKC is addedto each reaction to provide approximately 4 pmols phosphate transferredin 30 minute (per total reaction.) The reaction is linear over the timewhen 4 pmols of phosphate is transferred and the reaction remains linearwell beyond this time frame.

[0227] The PKC screening assay is performed in 96 well polystyrene Ubottom micro titer plates, in a total reaction volume of 50 ul. Solutionmanipulations are performed during the assay utilize a Rainin, motorizedEDP-plus M8 eight-channel micropipettor.

[0228] Samples were typically assayed at three dilutions, however somehighly active pure compounds were assayed at six dilutions. Assaysamples are dissolved in DMSO at a concentration of 10 mg/ml or less forsamples suspected of being more potent. In some cases 50% DMSO:water,water, or methanol is substituted (if essential) for the solvent. Atleast 25 μl of the highest concentration sample to be assayed istransferred to a well in a 96 well U-bottom polystyrene assay plate.Serial 5-fold or 10-fold dilutions (depending on the dose-range desired)are made using the EDP-plus M8 eight-channel pipettor in dilute mode andmixing by repipetting. Using the 8-channel pipettor, 2 μl of eachdilution is transferred to the appropriate wells of the plate(s) to beused for each assay. Duplicate assays are performed for each dose, witheach assay, allowing six wells (half the row) for three-dose assays, or12 wells (the whole row) for six-dose assays. In general, extracts andfractions are assayed at three doses: 400, 40 an 4 ug/ml, while purecompounds are tested at six doses: 400, 80, 16, 3.2, 0.64, 0.128 (8-foldseries). The results of these experiments are shown in Table 2. TABLE 2Inhibition of Protein Kinase C Compounds, NSC# Structure IC₅₀ (μg/ml)675347

1 × 10⁰ 675346

5 × 10⁻¹ 675345

9 × 10⁻¹ 675344

5 × 10¹ 675343

3 × 10¹ 676628

1 × 10⁰ 676629

2 × 10⁻¹ 676630

9 × 10⁻¹ 676631

2 × 10⁰ 676632

5 × 10¹ 676633

6 × 10⁰ 676634

8 × 10⁻¹ 676635

8 × 10⁰ 674973

7 × 10⁰ 675244

1 × 10⁰ 675245

1 × 10⁰ 675246

3 × 10⁰ 675247

1 × 10¹

1. A compound of formula I:

wherein R₁ and R₂ are independently selected form the group consistingof

 H, CH₂OH, CHO and CH₂NH₂; X and Y are independently selected from thegroup consisting of Se, S, O, and NR, wherein R is H or C₁-C₇ alkyl; R₃,R₄, R₅ and R₆ are independently selected from the group consisting of H,CHO, CH₂OH and CH₂NH₂; cyclodextrin complexes of such compounds; andwhen R₁, R₂, R₃, R₄, R₅ or R₆ is CH₂NH₂, the pharmaceutically acceptablesalt of the compound represented thereby; with the provisos, that R₁ andR₂ are not both

 and when R₁ and R₂ are both H, R₆ and R₃ are not both H; and  when R₂is

 one of R₁, R₃, R₄, R₅ and R₆ is other than H, and when R₁ is

 one of R₂, R₃, R₄, R₅ and R₆ is other than H.
 2. The compound of claim1, wherein R₃, R₄ and R₆ are H.
 3. The compound of claim 2 wherein R₂ isselected from the group consisting of H, CH₂OH, CHO and CH₂NH₂ and R₁ is


4. The compound of claim 2 wherein R₁ is selected from the groupconsisting of H, CH₂OH, CHO and CH₂NH₂ and R₂ is


5. The compound of claim 3 or 4 wherein X is Se.
 6. A compound offormula I:

wherein R₁ and R₂ are independently selected from the group consistingof

 H, CHO, CH₂OH and CH₂NH₂; X and Y are independently selected from thegroup consisting of Se, S, O and NR, wherein R is H or C₁-C₇ alkyl; R₃,R₄, R₅ and R₆ are independently selected from the group consisting of H,CHO, CH₂OH and CH₂NH₂; cyclodextrin complexes of such compounds; andwhen R₁, R₂, R₃, R₄, R₅ or R₆ is CH₂NH₂, the pharmaceutically acceptablesalt of the compound represented thereby; with the proviso that R₁ andR₂ are not both hydrogen, and when R₂ is

 R₁ is H, CHO, CH₂OH or CH₂NH₂, provided that at least one of R₁ R₃, R₄,R₅ and R₆ is other than H; and when R₁ is

 R₂ is H, CHO, CH₂OH or CH₂NH₂, provided that at least one of R₂, R₃,R₄, R₅ and R₆ is other than H.
 7. A composition comprising an anti-tumoreffective amount of a compound of formula I:

wherein R₁ and R₂ are independently selected from the group consistingof;

 H, CH₂OH, CHO and CH₂NH₂; X and Y are independently selected from thegroup consisting of Se, S, O and NR, wherein R is H or C₁-C₇ alkyl; R₃,R₄, R₅ and R₆ are independently selected from the group consisting of H,CHO, CH₂OH and CH₂NH₂; cyclodextrin complexes of such compounds; andwhen R₁, R₂, R₃, R₄, R₅ or R₆ is CH₂NH₂, the pharmaceutically acceptablesalt of the compound represented thereby; with the proviso, that R₁ andR₂ are not both

 and at least one of R₁, R₂, R₃, R₄, R₅ or R₆ is other than hydrogen;and a pharmaceutically acceptable carrier.
 8. The compound of claim 7,wherein R₃, R₄ and R₆ are H.
 9. The compound of claim 8 wherein R₂ isselected from the group consisting of H, CH₂OH, CHO and CH₂NH₂ and R₁ is


10. The compound of claim 8 wherein R₁ is selected from the groupconsisting of H, CH₂OH, CHO and CH₂NH₂ and R₂ is


11. The compound of claim 9 or 10 wherein X is Se.
 12. The use of acompound of the formula I:

wherein R₁ and R₂ are independently selected from the group consistingof;

 H, CHO, CH₂OH and CH₂NH₂; X and Y are independently selected from thegroup consisting of Se, S, O and NR, wherein R is H or C₁-C₇ alkyl; R₃,R₄, R₅ and R₆ are independently selected from the group consisting of H,CHO, CH₂OH and CH₂NH₂; cyclodextrin complexes of such compounds; andwhen R₃, R₄, R₅ or R₆ is CH₂NH₂, the pharmaceutically acceptable salt ofthe compound represented thereby; with the proviso, that R₁ and R₂ arenot both

to manufacture a pharmaceutical composition useful for treating apatient having a tumor.
 13. A method of preparing an intermediatecompound of the formula

wherin X and Y are selected from the group consisting of 0 Se, S and NR,wherein R is H or C₁-C₇ alkyl; and R₁, R₂, R₃, R₄ and R₆ areindependently selected from the group consisting of H, CHO, CH₂OH andCH₂NH₂, said method comprising the step of reacting a compound of theformula

 with a compound of the formula

 in the presence of sodium cyanide and in dimethyl formamide.
 14. Amethod of preparing a compound of the formula

wherein X, Y and Z are selected from the group consisting of O, Se, Sand NR, wherein R is H or C₁-C₇ alkyl; and R₁, R₂, R₃, R₄ and R₆ areindependently selected from the group consisting of H, CHO, CH₂OH andCH₂NH₂, said method comprising the steps of reacting a compound of theformula

 with a compound of the formula

 in the presence of sodium cyanide and DMF to form an intermediatehaving the formula

 and when Z is NR, reacting the intermediate with RNH₂Cl in the presenceof NaOAc;  when Z is O, reacting the intermediate with (CH₃CO)₂O in thepresence of HCl; and  when Z is S or Se, reacting the intermediate with[(C₆H₁₁)₃Sn]₂Z in the presence of BCl₃.